Hypersensitive reaction elicitor peptide and use thereof

Abstract

To provide a hypersensitive reaction induction peptide comprising improved solubility, stability, resistance to chemical decomposition, or combination thereof, for modulating a plant biochemical signal transmission, applying disease resistance to plants, enhancing plant growth, applying tolerance to biological stress, applying tolerance and resistance to non-biological stress, applying dryness resistance to a segmentation part cut from an ornamental plant, applying after-harvesting disease resistance or after-harvesting dryness resistance to fruit or vegetable, or extending a life of ripeness of fruit or vegetable.SOLUTION: There is provided an isolated peptide comprising amino acid length of less than 50 amino acids, and including a specific amino acid sequence.SELECTED DRAWING: None

Description

Technical Field

[0001] This application claims priority to U.S. Provisional Patent Application No. 62 / 058,535, filed Oct. 1, 2014, and U.S. Provisional Patent Application No. 62 / 140,789, filed Mar. 31, 2015, each of which is hereby incorporated by reference in its entirety.

[0002] Field of the Invention The present invention relates to novel hypersensitive response elicitor peptides and their use for inducing active plant responses, particularly including growth promotion, disease resistance, pest or insect resistance, and stress resistance.

Background Art

[0003] Background of the Invention The identification and isolation of harpin proteins originated from basic research at Cornell University that was attempting to understand how plant pathogenic bacteria interact with plants. The first line of defense is the hypersensitive response (HR), a localized plant cell death at the site of infection. The cell death creates a physical barrier against pathogen movement, and in some plants, the dead cells can release compounds toxic to the invading pathogen. Studies have shown that pathogenic bacteria likely have a single factor responsible for initiating HR. The basic objective of the research at Cornell University was to identify the specific bacterial protein responsible for inducing HR. The target protein was found to be encoded by one of a group of bacterial genes called the hypersensitive response and pathogenicity (hrp) gene cluster. The hrp cluster in Erwinia amylovora (Ea), the bacterium that causes fire blight in apple and pear, was analyzed, and a single protein that induces HR in certain plants was identified. This protein was named harpin (later, harpin EaIt was given the name ( ) and the corresponding gene was designated hrpN. This was the first example of such a protein and gene identified from any bacterial species.

[0004] Since then, several different harpin proteins have been identified, particularly from the genera Erwinia, Pseudomonas, Ralstonia, Xanthomonas, and Pantoea. Although diverse at the primary amino acid sequence level, harpin proteins share common biochemical and biophysical characteristics, as well as biological functions. Based on their unique properties, harpin proteins are considered in the literature to belong to a single group of proteins.

[0005] Following their identification and isolation, it was later discovered that harpins can induce disease resistance and enhance plant growth in plants. An important initial finding was that the application of purified harpin protein renders plants resistant to subsequent pathogen attacks and at plant locations quite distant from the injection site. This means that harpin proteins can trigger a plant defense mechanism called systemic acquired resistance (SAR), which confers resistance to various viral, bacterial, and fungal pathogens.

[0006] In crop protection, compositions that improve plant health are always in demand. Healthier plants are desirable as they result in better yields and / or better quality of plants or crops. Healthier plants are also better resistant to biotic and abiotic stresses. High resistance to biotic stresses in turn enables growers to reduce the amount of pesticides applied and, as a result, delay the development of resistance to individual pesticides.

[0007] Harpin αβ is a fusion protein derived from several different harpins. Harpin αβhas been found to suppress the production of nematode eggs, enhance plant growth, quality, and yield, and increase plant vigor. Its amino acid and nucleotide sequences are described in detail in US Patent Application Publication No. 2010 / 0043095 (Non-Patent Document 1).

[0008] To date, the production of harpins and their use in agricultural and horticultural applications have been as powdered solids coated on starch. This limits the use and versatility of harpin proteins, as liquid suspensions of powdered harpin proteins in water have a useful life of only 48 - 72 hours before significant loss of activity occurs. Another problem associated with harpin solutions is protein solubility and stability. αβ It would be desirable to identify synthetic and derivative harpin peptides that are highly soluble in aqueous solutions, stable, resistant to chemical degradation, and effective in initiating a hypersensitive response in plants.

[0009] It would be desirable to identify synthetic and derivative harpin peptides that are highly soluble in aqueous solutions, stable, resistant to chemical degradation, and effective in initiating a hypersensitive response in plants.

[0010] The present invention is directed to overcoming these and other limitations in the art.

Prior Art Documents

Non-Patent Documents

[0011]

Non-Patent Document 1

Summary of the Invention

[0012] A first aspect of the present invention is An isolated peptide having the amino acid sequence of TIFF2025096309000001.tif5163, wherein the peptide does not contain cysteine and methionine; each X at positions 2 and 6 is optional and, if present, can be any amino acid; and each X at positions 3, 7, 10, and 11 is any amino acid. In one embodiment, only one of the Xs at positions 2 and 6 is optional. In certain embodiments, SEQ ID NO:93 may further contain additional amino acid residues between the hydrophobic doublets (two L / I / V / F / A as shown). In certain embodiments, the isolated peptide further contains a hydrophilic amino acid sequence located at the N-terminus or C-terminus of SEQ ID NO:93.

[0013] A second aspect of the present invention is An isolated peptide having the amino acid sequence of TIFF2025096309000002.tif5163, wherein the peptide does not contain cysteine and methionine; each X at positions 2, 6, and 10 is optional and, if present, can be any amino acid; and each X at positions 3, 7, and 11 is any amino acid. In one embodiment, only one of the Xs at positions 2, 6, and 10 is optional. In certain embodiments, SEQ ID NO:93 may further contain additional amino acid residues between the hydrophobic doublets (two L / I / V / F / A as shown). In certain embodiments, the isolated peptide further contains a hydrophilic amino acid sequence located at the N-terminus or C-terminus of SEQ ID NO:93.

[0014] A third aspect of the present invention is Regarding an isolated peptide having the amino acid sequence of TIFF2025096309000003.tif4128 (SEQ ID NO:1, P1 / P4 consensus), wherein X at position 1 is optional and can be S, N, D, isoD, G, A, or S; X at position 2 is optional and can be Q, E, g-glutamate, G, A, or S; The 8-position X is Q, E, γ-glutamate, G, A, or S; The 9-position X is L, I, F, or V; The 10-position X is optional and can be D or iso-D; The 11-position X is Q, E, γ-glutamate, G, A, or S; The 12-position X is M, L, I, or F; The 13-position X is M, L, or I; The 14-position X is optional and can be any hydrophilic amino acid, preferably C, S, T, A, D, iso-D, K, or Q; The 15-position X is Q, E, γ-glutamate, G, A, S, K, or I; The 16-position X is M, L, I, V, or F; The 17-position X is M, L, I, A, or V; The 18-position X is Q, E, γ-glutamate, G, A, S, M, T, or K; The 19-position X is A, D, iso-D, S, V, T, K, R, E, H, or G; The 20-position X is M, L, or I; The 21-position X is M, L, I, V, S, or F; The 22-position X is Q, E, γ-glutamate, G, A, S; The 23-position X is P, Q, E, γ-glutamate, G, A, or S, and the isolated peptide contains one or more mutations compared to the corresponding wild-type amino acid sequence. In certain embodiments, the one or more mutations improve the water solubility, stability, or resistance to chemical degradation of the isolated peptide compared to a polypeptide containing the corresponding wild-type amino acid sequence.

[0015] One exemplary family of peptides according to the third aspect of the invention is has the amino acid sequence of TIFF2025096309000004.tif4128 (SEQ ID NO:2, P4 consensus), where The X at position 2 is Q, E, γ-glutamate, G, A, or S; The X at position 8 is Q, E, γ-glutamate, G, A, or S; The X at position 9 is L, A, D, isoD, I, V, or F; The X at position 11 is Q, E, γ-glutamate, G, A, or S; The X at position 12 is L, D, isoD, I, or F; The X at position 13 is L, I, V, or F; The X at position 14 is any hydrophilic amino acid, preferably C, S, or T, S or T, or S only; The X at position 15 is Q, E, γ-glutamate, G, A, S, K, or I; The X at position 16 is L, A, I, V, M, or F; The X at position 17 is I, S, or F; The X at position 18 is Q, E, γ-glutamate, G, A, or S; The X at position 20 is L, I, V, or F; The X at position 21 is L or F; and The X at position 22 is Q, E, γ-glutamate, G, A, or S. In certain embodiments, these peptides according to the third aspect of the invention also have the structural features that define the peptides according to the first or second aspect of the invention.

[0016] Another exemplary family of peptides according to the third aspect of the invention is having the amino acid sequence of TIFF2025096309000005.tif4128 (SEQ ID NO:3, P1 consensus), where The X at position 1 is N, D, isoD, G, A, or S; The X at position 2 is Q, E, γ-glutamate, G, A, or S; The X at position 8 is Q, E, γ-glutamate, G, A, or S; The X at position 11 is Q, E, γ-glutamate, G, A, or S; The 15-position X is Q, E, γ-glutamate, G, A, or S; The 18-position X is M, T, K, E, γ-glutamate, G, A, or S; The 22-position X is Q, E, γ-glutamate, G, A, or S; and The 23-position X is Q, E, γ-glutamate, G, A, or S. In certain embodiments, these peptides according to the third aspect of the invention also have the structural features that define the peptides according to the first or second aspect of the invention.

[0017] The fourth aspect of the invention is (i) the 3-position X is N, D, or isoD; the 6-position X is Q, E, γ-glutamate, G, A, or S; the 8-position X is N, D, or isoD; the 15-position X is optional and can be any amino acid; the 18-position X is M, E, γ-glutamate, G, A, S, T, or K; and the 22-position X is optional and can be Q, E, γ-glutamate, G, A, or S, TIFF2025096309000006.tif4128 (SEQ ID NO:47, P15b / P20 consensus), or (ii) the 7-position X is optional and can be any amino acid; the 10-position X is M, E, γ-glutamate, G, A, S, T, or K; and the 14-position X is optional and can be Q, E, γ-glutamate, G, A, or S, TIFF2025096309000007.tif4128 (SEQ ID NO:12, P15 / 20min consensus) Relates to an isolated peptide having the amino acid sequence. In certain embodiments, the isolated peptide contains one or more mutations compared to the corresponding wild-type amino acid sequence, and one or more of those mutations improve the water solubility, stability, or resistance to chemical degradation of the isolated peptide compared to the polypeptide containing the corresponding wild-type amino acid sequence. In certain embodiments, the peptide according to the fourth aspect of the present invention also has the structural features defining the peptide according to the first or second aspect of the present invention.

[0018] The fifth aspect of the present invention is (i) X at position 4 is F or Y; X at position 6 is Q, E, γ-glutamate, G, A, or S; X at position 7 is optional and can be L, M, E, γ-glutamate, G, A, S, T, or K; X at position 9 is M, E, γ-glutamate, G, A, S, T, or K; X at position 10 is H or N; X at position 14 is E, γ-glutamate, D, or isoD; X at position 17 is Q, E, γ-glutamate, G, A, or S; and X at position 19 is Q, E, γ-glutamate, G, A, or S, TIFF2025096309000008.tif4128 (SEQ ID NO:66, P6 / 6a consensus), or (ii) X at position 1 is F or Y; X at position 3 is Q, E, γ-glutamate, G, A, or S; X at position 4 is optional and, according to one embodiment, can be M, E, γ-glutamate, G, A, S, T, or K, or according to another embodiment, can be L; X at position 6 is M, E, γ-glutamate, G, A, S, T, or K; X at position 7 is H or N; and wherein the 11 - position X is E, γ - glutamate, D, or iso - D, TIFF2025096309000009.tif4128 (SEQ ID NO:135, P6 / 6a min consensus) having the amino acid sequence of , containing one or more mutations as compared with the corresponding wild - type amino acid sequence, and wherein said one or more mutations improve the water solubility, stability, or resistance to chemical degradation of the isolated peptide as compared with the polypeptide containing the corresponding wild - type amino acid sequence. In certain embodiments, the peptide according to the fifth aspect of the invention also has the structural features that define the peptide according to the first or second aspect of the invention.

[0019] The sixth aspect of the invention (i) the 1 - position X can be Q, N, D, E, γ - glutamate, iso - D, or S; the 2 - position X can be D, E, γ - glutamate, iso - D; the 3 - position X can be P, D, E, iso - D, or γ - glutamate; the 4 - position X can be M, A, S, D, E, iso - D, or γ - glutamate; the 5 - position X can be Q, E, or γ - glutamate; the 6 - position X can be A, E, or γ - glutamate; the 8 - position X can be M, L, E, Q, D, N, G, A, S, iso - D, or γ - glutamate; the 9 - position X can be Q, N, E, D, G, A, S, iso - D, or γ - glutamate; the 12 - position X can be Q, N, E, D, G, A, S, iso - D, or γ - glutamate; the 13 - position X can be Q, N, E, D, G, A, S, iso - D, or γ - glutamate; and the 16 - position X can be K, Q, N, E, D, R, G, A, or S, TIFF2025096309000010.tif4128 (SEQ ID NO:13, P14d consensus), or (ii) The two Xs can be M, L, E, Q, D, N, G, A, S, isoD, or g-glutamate; The three X can be Q, N, E, D, G, A, S, isoD, or g-glutamate; The six X can be Q, N, E, D, G, A, S, isoD, or g-glutamate; The seven X can be Q, N, E, D, G, A, S, isoD, or g-glutamate; and The ten X can be K, Q, N, E, D, R, G, A, or S, TIFF2025096309000011.tif4128 (SEQ ID NO:14, P14d min consensus) relates to an isolated peptide having the amino acid sequence of. In certain embodiments, the isolated peptide contains one or more mutations compared to the corresponding wild-type amino acid sequence, and one or more of those mutations improve the water solubility, stability, or resistance to chemical degradation of the isolated peptide compared to the polypeptide containing the corresponding wild-type amino acid sequence. In certain embodiments, the peptide according to the sixth aspect of the invention also has the structural features defining the peptide according to the first or second aspect of the invention.

[0020] The seventh aspect of the invention is (i) The two X can be Q, N, E, g-glutamate, D, isoD, T, S, A, or G; The three X can be K, Q, N, E, g-glutamate, D, isoD, T, S, A, or G; The six X can be K, Q, N, E, g-glutamate, D, isoD, T, S, A, or G; The nine X can be E, g-glutamate, D, isoD, Q, N, T, S, A, or G; and The ten X can be A, G, S, T, E, g-glutamate, D, isoD, Q, or N, TIFF2025096309000012.tif5128 (SEQ ID NO:16, P25 consensus), or (ii) The two Xs can be T, S, A, G, D, isoD, E, g-glutamate, Q, or N; The three Xs can be G, T, S, A, D, isoD, E, g-glutamate, Q, or N; The six Xs can be Q, N, E, g-glutamate, D, isoD, T, S, A, or G; The seven Xs can be K, Q, N, E, g-glutamate, D, isoD, T, S, A, or G; The ten Xs can be K, Q, N, E, g-glutamate, D, isoD, T, S, A, or G; The thirteen Xs can be E, g-glutamate, D, isoD, Q, N, T, S, A, or G; The fourteen Xs can be A, G, S, T, E, g-glutamate, D, isoD, Q, or N; and The sixteen V is optional, TIFF2025096309000013.tif5128 (SEQ ID NO:17, P25 consensus) relates to an isolated peptide having the amino acid sequence. In certain embodiments, the isolated peptide contains one or more mutations compared to the corresponding wild-type amino acid sequence, and one or more of those mutations improve the water solubility, stability, or resistance to chemical degradation of the isolated peptide compared to the polypeptide containing the corresponding wild-type amino acid sequence. In certain embodiments, the peptide according to the seventh aspect of the present invention also has the structural features defining the peptide according to the first or second aspect of the present invention.

[0021] The eighth aspect of the present invention is (i) The one X can be any amino acid, provided that preferably it can be Q, S, E, g-glutamate, A, T, G, D, isoD, N, K, or R; The two Xs can be any amino acid, provided that preferably it can be Q, S, E, g-glutamate, A, T, G, D, isoD, N, K, or R; The X at the 3rd position can be any amino acid, but preferably P, Q, S, E, γ-glutamate, A, T, G, D, iso-D, N, K, or R; The X at the 4th position can be any amino acid, but preferably I, Q, S, E, γ-glutamate, A, T, G, D, N, iso-D, K, or R; The X at the 5th position can be any amino acid, but preferably D, iso-D, S, E, γ-glutamate, A, T, G, N, Q, K, or R; The X at the 6th position can be any amino acid, but preferably R, Q, S, E, γ-glutamate, A, T, G, D, iso-D, N, or K; The X at the 7th position can be any amino acid, but preferably Q, S, E, γ-glutamate, A, T, G, D, iso-D, N, K, or R; The X at the 8th position can be any amino acid, but preferably T, Q, S, E, γ-glutamate, A, G, D, iso-D, N, K, or R; The X at the 9th position can be any amino acid, but preferably I, Q, S, E, γ-glutamate, A, T, G, D, iso-D, N, K, or R; The X at the 10th position can be any amino acid, but preferably E, γ-glutamate, Q, S, A, T, G, D, iso-D, N, K, or R; The X at the 11th position can be any amino acid, but preferably Q, S, E, γ-glutamate, A, T, G, D, iso-D, N, K, or R; The X at the 13th position can be any amino acid, but preferably A, S, T, G, D, iso-D, E, γ-glutamate, Q, N, K, or R; The X at the 14th position can be any amino acid, but preferably Q, A, S, T, G, D, iso-D, E, γ-glutamate, N, K, or R; The X at the 17th position can be any amino acid, but preferably A, S, T, G, D, iso-D, E, γ-glutamate, Q, N, K, or R; The X at the 18th position can be any amino acid, but preferably Q, A, S, T, G, D, iso-D, E, γ-glutamate, N, K, or R; The X at position 21 can be any amino acid, but preferably K, A, S, T, G, D, isoD, E, γ-glutamate, Q, N, or R; The X at position 22 can be any amino acid, but preferably S, A, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; The X at position 25 can be any amino acid, but preferably S, A, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; The X at position 26 can be any amino acid, but preferably P, S, A, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; and The X at position 27 can be any amino acid, but preferably Q, S, A, T, G, D, isoD, E, γ-glutamate, N, K, or R, TIFF2025096309000014.tif5128 (SEQ ID NO:21, P17 / 18), or (ii) The X at position 2 can be any amino acid, but preferably A, S, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; The X at position 3 can be any amino acid, but preferably Q, A, S, T, G, D, isoD, E, γ-glutamate, N, K, or R; The X at position 6 can be any amino acid, but preferably A, S, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; The X at position 7 can be any amino acid, but preferably Q, A, S, T, G, D, isoD, E, γ-glutamate, N, K, or R; The X at position 10 can be any amino acid, but preferably K, A, S, T, G, D, isoD, E, γ-glutamate, Q, N, or R; and The X at position 11 can be any amino acid, but preferably S, A, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R, TIFF2025096309000015.tif5128 (SEQ ID NO:25, P17 / 18min consensus) having the amino acid sequence of , including one or more mutations as compared to the corresponding wild-type amino acid sequence, and the one or more mutations improve the water solubility, stability, or resistance to chemical degradation of the isolated peptide as compared to the polypeptide including the corresponding wild-type amino acid sequence, relating to an isolated peptide. In certain embodiments, the peptide according to the eighth aspect of the present invention also has the structural features defining the peptide according to the first or second aspect of the present invention.

[0022] The ninth aspect of the present invention is X at position 1 is optional and can be L, I, V, F, or M; X at position 3 is any amino acid, provided that it can preferably be K, A, S, T, G, D, isoD, E, γ-glutamate, Q, N, or R; X at position 4 is any amino acid, provided that it can preferably be A, S, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; X at position 7 is any amino acid, provided that it can preferably be K, A, S, T, G, D, isoD, E, γ-glutamate, Q, N, or R; X at position 10 is any amino acid, provided that it can preferably be A, S, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; and X at position 11 is any amino acid, provided that it can preferably be R, A, S, T, G, D, isoD, E, γ-glutamate, Q, N, or K, relating to an isolated peptide having the amino acid sequence of TIFF2025096309000016.tif5128 (SEQ ID NO:26, P19 consensus). In certain embodiments, the isolated peptide includes one or more mutations as compared to the corresponding wild-type amino acid sequence, and the one or more mutations improve the water solubility, stability, or resistance to chemical degradation of the isolated peptide as compared to the polypeptide including the corresponding wild-type amino acid sequence. In certain embodiments, the peptide according to the ninth aspect of the present invention also has the structural features defining the peptide according to the first or second aspect of the present invention.

[0023] The tenth aspect of the present invention is that the X at the 2nd position can be Q, N, E, g-glutamate, D, isoD, T, S, A, or G; the X at the 3rd position can be Q, N, E, g-glutamate, D, isoD, T, S, A, or G; the X at the 6th position can be K, Q, N, E, g-glutamate, D, isoD, T, S, A, or G; the X at the 9th position can be E, g-glutamate, D, isoD, Q, N, T, S, A, or G; the X at the 10th position can be A, G, S, T, E, g-glutamate, D, isoD, Q, or N; the X at the 13th position can be Q, N, E, g-glutamate, D, isoD, T, S, A, or G; and the X at the 14th position can be Q, N, E, g-glutamate, D, isoD, T, S, A, or G, relates to an isolated peptide comprising the amino acid sequence of TIFF2025096309000017.tif5128 (SEQ ID NO:15, P3min consensus). In certain embodiments, the isolated peptide comprises one or more mutations compared to the corresponding wild-type amino acid sequence, and one or more of those mutations improve the water solubility, stability, or resistance to chemical degradation of the isolated peptide compared to a polypeptide comprising the corresponding wild-type amino acid sequence. In certain embodiments, the peptide according to the tenth aspect of the present invention also has the structural features defining the peptide according to the first or second aspect of the present invention.

[0024] The eleventh aspect of the present invention relates to a fusion protein comprising one of the peptides of the first to eleventh aspects of the present invention together with a purification tag, a solubility tag, or one or more of the second peptides according to one of the first to tenth aspects of the present invention.

[0025] The twelfth aspect of the present invention relates to a composition comprising one or more peptides according to the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, or tenth aspect of the present invention or a fusion protein according to the eleventh aspect of the present invention and a carrier.

[0026] The thirteenth aspect of the present invention relates to a method for conferring disease resistance to a plant. This method includes applying an effective amount of an isolated peptide according to the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, or tenth aspect of the present invention, a fusion protein according to the eleventh aspect of the present invention, or a composition according to the twelfth aspect of the present invention to a plant or a plant seed or a place where the plant is growing or is predicted to grow, and the application is effective for conferring disease resistance.

[0027] The fourteenth aspect of the present invention relates to a method for increasing plant growth. This method includes applying an effective amount of an isolated peptide according to the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, or tenth aspect of the present invention, a fusion protein according to the eleventh aspect of the present invention, or a composition according to the twelfth aspect of the present invention to a plant or a plant seed or a place where the plant is growing or is predicted to grow, and the application is effective for promoting plant growth.

[0028] The fifteenth aspect of the present invention relates to a method for increasing the tolerance and resistance of a plant to biotic stressors. This method includes applying an effective amount of an isolated peptide according to the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, or tenth aspect of the present invention, a fusion protein according to the eleventh aspect of the present invention, or a composition according to the twelfth aspect of the present invention to a plant or a plant seed or a place where the plant is growing or is predicted to grow, and the application is effective for increasing the tolerance and resistance of the plant to biotic stressors selected from the group consisting of pests such as insects, spiders, nematodes, weeds, and combinations thereof.

[0029] The sixteenth aspect of the present invention relates to a method for increasing the tolerance of plants to abiotic stress. This method comprises applying an effective amount of an isolated peptide according to the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, or tenth aspect of the present invention, a fusion protein according to the eleventh aspect of the present invention, or a composition according to the twelfth aspect of the present invention to a plant or a plant seed or to a location where a plant is growing or is predicted to grow, said application being effective for increasing the tolerance of the plant to an abiotic stress factor selected from the group consisting of salt stress, water stress (including drought and irrigation), ozone stress, heavy metal stress, low temperature stress, high temperature stress, nutrient stress (phosphate, potassium, nitrogen deficiency), bleaching and photoinduced stress, and combinations thereof.

[0030] The seventeenth aspect of the present invention relates to a method for imparting drought resistance to an excised portion taken from an ornamental plant. This method comprises applying an isolated peptide according to the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, or tenth aspect of the present invention, a fusion protein according to the eleventh aspect of the present invention, or a composition according to the twelfth aspect of the present invention to a plant or to a location where a plant is growing, said application being effective for imparting drought resistance to an excised portion taken from an ornamental plant.

[0031] The eighteenth aspect of the present invention relates to a method for imparting post-harvest disease resistance or post-harvest drought resistance to fruits or vegetables. This method comprises applying an effective amount of an isolated peptide according to the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, or tenth aspect of the present invention, a fusion protein according to the eleventh aspect of the present invention, or a composition according to the twelfth aspect of the present invention to a plant including the fruit or vegetable or to a location where the plant is growing; or applying an effective amount of the isolated peptide or composition thereto to the harvested fruit or vegetable, said application being effective for imparting post-harvest disease resistance or post-harvest drought resistance to the fruit or vegetable.

[0032] The 19th aspect of the present invention relates to a method for extending the shelf life of fruits or vegetables. This method includes applying an effective amount of an isolated peptide according to the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, or 10th aspect of the present invention, a fusion protein according to the 11th aspect of the present invention, or a composition according to the 12th aspect of the present invention to a plant containing fruits or vegetables, or to the location where the plant is growing; or applying an effective amount of the isolated peptide or the composition thereof to the harvested fruits or vegetables, and the application is effective for extending the shelf life of the fruits or vegetables.

[0033] The 20th aspect of the present invention relates to a method for modulating one or more biological signal transduction processes in plants. This method includes applying an effective amount of an isolated peptide according to the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, or 10th aspect of the present invention, a fusion protein according to the 11th aspect of the present invention, or a composition according to the 12th aspect of the present invention to a plant or to the location where the plant is growing, and the application is effective for modulating one or more biochemical signal transduction processes.

[0034] The 21st aspect of the present invention relates to a DNA construct comprising a first nucleic acid molecule encoding a polypeptide according to the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, or 10th aspect of the present invention or a fusion protein according to the 11th aspect of the present invention; and a nucleic acid molecule effective as a promoter operably coupled to the first nucleic acid molecule. This aspect of the present invention also includes a recombinant expression vector containing the DNA construct, a recombinant host cell containing the DNA construct, and further includes a transgenic plant or plant seed containing the recombinant plant cell of the present invention (containing the DNA construct).

[0035] The 22nd aspect of the present invention relates to a method of conferring disease resistance to a plant, promoting plant growth, conferring tolerance and resistance to biotic stressors, conferring tolerance to abiotic stress, or modulating plant biochemical signal transduction. This method comprises providing a transformed plant transformed with a DNA construct according to the 21st aspect of the present invention; and the DNA construct expressing the peptide or the fusion polypeptide to confer disease resistance to the transformed plant, to enable promotion of plant growth, to enable conferring tolerance to biotic stress, to enable conferring tolerance to abiotic stress, or to enable modulating biochemical signal transduction, growing the plant under conditions effective therefor.

[0036] The 23rd aspect of the present invention relates to a method of conferring drought resistance to an excised portion taken from an ornamental plant, conferring post-harvest disease resistance or post-harvest drought resistance to a fruit or vegetable, or extending the shelf life of a fruit or vegetable. The method comprises providing a transformed plant transformed with a DNA construct comprising a first nucleic acid molecule encoding a polypeptide according to the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, 9th, or 10th aspect of the present invention or a fusion protein according to the 11th aspect of the present invention; and the DNA construct expressing the peptide or the fusion polypeptide to confer drought resistance to an excised portion taken from the transformed ornamental plant, to enable conferring post-harvest disease resistance or post-harvest drought resistance to a fruit or vegetable taken from the transformed plant, or to enable extending the shelf life of a fruit or vegetable taken from the transformed plant, growing the plant under conditions effective therefor.

[0037] The 24th aspect of the present invention relates to a method of conferring disease resistance to plants, promoting plant growth, conferring tolerance and resistance to biotic stressors, conferring tolerance to abiotic stress, or modulating biochemical signal transduction. This method comprises providing a transformed plant seed transformed with a DNA construct according to the 21st aspect of the present invention; planting the transformed plant seed in soil; and propagating a transformed plant from the transformed plant seed in order for the DNA construct to express the peptide or the fusion polypeptide to confer disease resistance, to enable promotion of plant growth, to enable conferring tolerance to biotic stress, or to enable conferring tolerance to abiotic stress.

[0038] The 25th aspect of the present invention relates to a method of conferring drought resistance to an excised portion taken from an ornamental plant, conferring post-harvest disease resistance or post-harvest drought resistance to a fruit or vegetable, or extending the shelf life of a fruit or vegetable. The method comprises providing a transformed plant seed transformed with a DNA construct according to the 21st aspect of the present invention; planting the transformed plant seed in soil; and propagating the transformed plant from the transformed plant seed in order for the DNA construct to express the peptide or the fusion polypeptide to enable conferring drought resistance to an excised portion taken from the transformed ornamental plant, to enable conferring post-harvest disease resistance or post-harvest drought resistance to a fruit or vegetable taken from the transformed plant, or to enable extending the shelf life of a fruit or vegetable taken from the transformed plant.

[0039] By providing HR-inducing peptides that exhibit improved solubility, stability, resistance to chemical degradation, or combinations of these properties, this will give growers greater flexibility in the preparation, handling, and delivery of effective amounts of compositions containing these HR-inducing peptides to plants in the field or greenhouse. For growers, simplification of the application process leads to greater compliance and thus improved results with respect to one or more of disease resistance, growth promotion, tolerance and resistance to biotic stressors, tolerance to abiotic stress, resistance to drying in excised portions taken from ornamental plants, post-harvest disease resistance or post-harvest drying resistance of fruits or vegetables harvested from plants, and / or improvement in the shelf life of fruit or vegetable ripening for fruits or vegetables harvested from plants. These and other advantages are described herein. [Inventive Concept 1001] An isolated peptide comprising the amino acid sequence of TIFF2025096309000018.tif5164, wherein it does not contain cysteine and methionine; each X at positions 2, 6, and 10 is optional and, if present, is any amino acid; and each X at positions 3, 7, and 11 is any amino acid. [Inventive Concept 1002] The isolated peptide of Inventive Concept 1001 that is less than 100 amino acids in length. [Inventive Concept 1003] The isolated peptide of Inventive Concept 1002 that is 13 to 50 amino acids in length. [Inventive Concept 1004] The isolated peptide of Inventive Concept 1001 that is stable when dissolved in water or an aqueous solution. [Inventive Concept 1005] The isolated peptide of Inventive Concept 1001 that is resistant to chemical degradation when dissolved in an aqueous buffer containing a biocide. [Inventive Concept 1006] An isolated peptide of the present invention 1001 having a solubility of more than about 0.1% in water or an aqueous solution. [The present invention 1007] An isolated peptide of the present invention 1001 in which one or both of the Xs at positions 2 and 6 are absent. [The present invention 1008] An isolated peptide of the present invention 1001 in which both of the Xs at positions 2 and 6 are present. [The present invention 1009] An isolated peptide of the present invention 1001 or 1007 in which the X at position 10 is absent. [The present invention 1010] An isolated peptide of the present invention 1001 or 1008 in which the X at position 10 is present. [The present invention 1011] Each X at positions 2 and 6, when present, is a polar or charged amino acid; and An isolated peptide of the present invention 1001 in which each X at positions 3, 7, 10, and 11 is a polar or charged amino acid. [The present invention 1012] Each X at positions 2 and 6, when present, is selected from the group consisting of R, K, D, isoD, E, N, Q, H, S, T, Y, W, G, A, and g-glutamate; and An isolated peptide of the present invention 1001 in which each X at positions 3, 7, 10, and 11 is selected from the group consisting of R, K, D, isoD, E, N, Q, H, S, T, Y, W, G, A, and g-glutamate. [The present invention 1013] Each X at positions 2 and 6, when present, is selected from the group consisting of D, isoD, E, and g-glutamate; and An isolated peptide of the present invention 1001 in which each X at positions 3, 7, 10, and 11 is selected from the group consisting of D, isoD, E, and g-glutamate. [The present invention 1014] An isolated peptide of the present invention 1013 comprising the amino acid sequence of TIFF2025096309000019.tif92128. [The present invention 1015] The isolated peptide of the present invention 1001, further comprising a hydrophilic amino acid sequence at either the N-terminus or the C-terminus of SEQ ID NO:93. [The present invention 1016] The isolated peptide of the present invention 1015, comprising one of the amino acid sequences of SEQ ID NO:31, 33, 126, 134, 141, 149, 150, 161, 166, 167, 168 - 173, 178, 187 - 189, 200 - 202, 206 - 209, 231, or 232. [The present invention 1017] The isolated peptide of the present invention 1001, comprising the amino acid sequence of TIFF2025096309000020.tif238133. [The present invention 1018] Comprising the amino acid sequence of TIFF2025096309000021.tif4128 (SEQ ID NO:1, P1 / P4 consensus), where X at position 1 is optional and can be S, N, D, isoD, G, A, or S; X at position 2 is optional and can be Q, E, g-glutamate, G, A, or S; X at position 8 is Q, E, g-glutamate, G, A, or S; X at position 9 is L, I, F, or V; X at position 10 is optional and can be D or isoD; X at position 11 is Q, E, g-glutamate, G, A, or S; X at position 12 is M, L, I, or F; X at position 13 is M, L, or I; X at position 14 is optional and can be any hydrophilic amino acid; X at position 15 is Q, E, g-glutamate, G, A, S, K, or I; X at position 16 is M, L, I, V, or F; X at position 17 is M, L, I, A, or V; X at position 18 is Q, E, g-glutamate, G, A, S, M, T, or K; The 19th X is A, D, isoD, S, V, T, K, R, E, H, or G; The 20th X is M, L, or I; The 21st X is M, L, I, V, S, or F; The 22nd X is Q, E, g-glutamate, G, A, S; The 23rd X is P, Q, E, g-glutamate, G, A, or S, and The isolated peptide contains one or more mutations as compared to the corresponding wild-type amino acid sequence, and the one or more mutations improve the stability or resistance to chemical degradation of the isolated peptide as compared to a polypeptide containing the corresponding wild-type amino acid sequence, said isolated peptide. [Inventive item 1019] The isolated peptide of Inventive item 1018, which does not consist of TIFF2025096309000022.tif5128 (P1, SEQ ID NO:4). [Inventive item 1020] The isolated peptide of Inventive item 1018, which does not consist of TIFF2025096309000023.tif5128 (P4, SEQ ID NO:5). [Inventive item 1021] The isolated peptide of Inventive item 1018, wherein the polypeptide containing the corresponding wild-type amino acid sequence consists of SEQ ID NO:4 or 5, and the isolated peptide is more stable than the polypeptide of SEQ ID NO:4 or 5 when dissolved in water or an aqueous solution. [Inventive item 1022] The isolated peptide of Inventive item 1018, wherein the polypeptide containing the corresponding wild-type amino acid sequence is SEQ ID NO:4 or 5, and the isolated peptide has higher resistance to chemical degradation than the polypeptide of SEQ ID NO:4 or 5 when dissolved in an aqueous buffer containing a biocide. [Inventive item 1023] TIFF2025096309000024.tif4128 (SEQ ID NO:2, P4 consensus) contains the amino acid sequence, where X at position 2 is Q, E, γ-glutamate, G, A, or S; X at position 8 is Q, E, γ-glutamate, G, A, or S; X at position 9 is L, A, D, isoD, I, V, or F; X at position 11 is Q, E, γ-glutamate, G, A, or S; X at position 12 is L, D, isoD, I, or F; X at position 13 is L, I, V, or F; X at position 14 is any hydrophilic amino acid; X at position 15 is Q, E, γ-glutamate, G, A, S, K, or I; X at position 16 is L, A, I, V, M, or F; X at position 17 is I, S, or F; X at position 18 is Q, E, γ-glutamate, G, A, or S; X at position 20 is L, I, V, or F; X at position 21 is L or F; and X at position 22 is Q, E, γ-glutamate, G, A, or S, the isolated peptide of the present invention 1018. [The present invention 1024] The isolated peptide of the present invention 1023, where X at position 14 is S. [The present invention 1025] The isolated peptide of the present invention 1023, containing one of the amino acid sequences of SEQ ID NO:6 - 11, 19, 20, 22 - 24, 27 - 30, 32, 34, 37, 40, 98 - 108, 111 - 114, 116, 117, 119 - 125, 127 - 131, 136 - 139, 191, and 196. [The present invention 1026] TIFF2025096309000025.tif4128 (SEQ ID NO:3, P1 consensus) contains the amino acid sequence, where The X at position 1 is N, D, isoD, G, A, or S; The X at position 2 is Q, E, g-glutamate, G, A, or S; The X at position 8 is Q, E, g-glutamate, G, A, or S; The X at position 11 is Q, E, g-glutamate, G, A, or S; The X at position 15 is Q, E, g-glutamate, G, A, or S; The X at position 18 is M, T, K, E, g-glutamate, G, A, or S; The X at position 22 is Q, E, g-glutamate, G, A, or S; and The X at position 23 is Q, E, g-glutamate, G, A, or S, an isolated peptide of the present invention 1018. [The present invention 1027] For at least one of positions 2, 8, 11, 15, 22, and 23 of SEQ ID NO:3, X is E, g-glutamate, G, A, or S, an isolated peptide of the present invention 1026. [The present invention 1028] An isolated peptide of the present invention 1026, comprising one of the amino acid sequences of SEQ ID NOs: 41 to 46, 109, 110, 115, and 118. [The present invention 1029] An isolated peptide of the present invention 1018, having a length of less than 100 amino acids. [The present invention 1030] An isolated peptide of the present invention 1029, having a peptide length of 23 to 50. [The present invention 1031] An isolated peptide of the present invention 1018, consisting essentially of the amino acid sequence described. [The present invention 1032] An isolated peptide of the present invention 1018, consisting of the amino acid sequence described. [The present invention 1033] An isolated peptide having a length of less than 100 amino acids and comprising the amino acid sequence of SEQ ID NO:5. [The present invention 1034] An isolated peptide of the invention 1033 consisting essentially of SEQ ID NO:5. [Invention 1035] An isolated peptide of the invention 1033 consisting of SEQ ID NO:5. [Invention 1036] (i) X at position 3 is N, D, or isoD; X at position 6 is Q, E, g-glutamate, G, A, or S; X at position 8 is N, D, or isoD; X at position 15 is optional and can be any amino acid; X at position 18 is M, E, g-glutamate, G, A, S, T, or K; and X at position 22 is optional and can be Q, E, g-glutamate, G, A, or S, TIFF2025096309000026.tif4128 (SEQ ID NO:47, P15b / P20 consensus), or (ii) X at position 7 is optional and can be any amino acid; X at position 10 is M, E, g-glutamate, G, A, S, T, or K; and X at position 14 is optional and can be Q, E, g-glutamate, G, A, or S, TIFF2025096309000027.tif4128 (SEQ ID NO:12, P15 / 20min consensus) comprising the amino acid sequence of, containing one or more mutations as compared to the corresponding wild-type amino acid sequence, said one or more mutations improving the stability or resistance to chemical degradation of the isolated peptide as compared to a polypeptide comprising the corresponding wild-type amino acid sequence, said isolated peptide. [Invention 1037] An isolated peptide of the invention 1036 having a length of less than 100 amino acids. [Invention 1038] The polypeptide comprising the corresponding wild-type amino acid sequence is SEQ ID NO:48, and the isolated peptide is more stable than the polypeptide of SEQ ID NO:48 when dissolved in water or an aqueous solution. The isolated peptide of the present invention 1036. [The present invention 1039] The polypeptide comprising the corresponding wild-type amino acid sequence is SEQ ID NO:48, and the isolated peptide has higher resistance to chemical degradation than the polypeptide of SEQ ID NO:48 when dissolved in an aqueous buffer containing a biocide. The isolated peptide of the present invention 1036. [The present invention 1040] The isolated peptide of the present invention 1036, comprising one amino acid sequence selected from SEQ ID NOs: 49 to 65, 142 to 144, 151, or 152. [The present invention 1041] The isolated peptide of the present invention 1036, consisting essentially of the amino acid sequence described. [The present invention 1042] The isolated peptide of the present invention 1036, consisting of the amino acid sequence described. [The present invention 1043] An isolated peptide comprising 22 to 36 amino acids, consisting essentially of the amino acids of SEQ ID NO:49, SEQ ID NO:63, or SEQ ID NO:64. [The present invention 1044] An isolated peptide comprising the amino acid sequence of SEQ ID NO:65. [The present invention 1045] (i) X at position 4 is F or Y; X at position 6 is Q, E, γ-glutamate, G, A, or S; X at position 7 is optional and can be M, E, γ-glutamate, G, A, S, T, or K; X at position 9 is M, E, γ-glutamate, G, A, S, T, or K; X at position 10 is H or N; The 14-position X is E, γ-glutamate, D, or isoD; The 17-position X is Q, E, γ-glutamate, G, A, or S; and The 19-position X is Q, E, γ-glutamate, G, A, or S, TIFF2025096309000028.tif4128 (SEQ ID NO:66, peptide 6 / 6a consensus), or (ii) The 1-position X is F or Y; The 3-position X is Q, E, γ-glutamate, G, A, or S; The 4-position X is optional and, according to one embodiment, can be M, E, γ-glutamate, G, A, S, T, or K, or according to another embodiment, can be L; The 6-position X is M, E, γ-glutamate, G, A, S, T, or K; The 7-position X is H or N; and The 11-position X is E, γ-glutamate, D, or isoD, TIFF2025096309000029.tif4128 (SEQ ID NO:135, P6 / 6a min consensus) comprising the amino acid sequence of, containing one or more mutations as compared to the corresponding wild-type amino acid sequence, wherein the one or more mutations improve the stability or resistance to chemical degradation of the isolated peptide as compared to the polypeptide comprising the corresponding wild-type amino acid sequence, said isolated peptide. [Inventive concept 1046] The isolated peptide of inventive concept 1045, which does not consist of TIFF2025096309000030.tif5128 (P6a, SEQ ID NO:67). [Inventive concept 1047] The isolated peptide of inventive concept 1045, wherein the polypeptide comprising the corresponding wild-type amino acid sequence is SEQ ID NO:67, and the isolated peptide is more stable than the polypeptide of SEQ ID NO:67 when dissolved in water or an aqueous solution. [The present invention 1048] The polypeptide comprising the corresponding wild-type amino acid sequence is SEQ ID NO:67, and the isolated peptide has higher resistance to chemical degradation than the polypeptide of SEQ ID NO:67 when dissolved in an aqueous buffer containing a biocide, the isolated peptide of the present invention 1045. [The present invention 1049] The isolated peptide of the present invention 1045, comprising one amino acid sequence among SEQ ID NO:68 - 80, 155 - 161, 197, and 198. [The present invention 1050] The isolated peptide of the present invention 1045, having less than 100 amino acids in length. [The present invention 1051] The isolated peptide of the present invention 1045, having a peptide length of 20 - 50. [The present invention 1052] The isolated peptide of the present invention 1045, consisting essentially of the recited amino acid sequence. [The present invention 1053] The isolated peptide of the present invention 1045, consisting of the recited amino acid sequence. [The present invention 1054] The isolated peptide of the present invention 1045, not including SEQ ID NO:154. [The present invention 1055] The isolated peptide of the present invention 1045, comprising the amino acid sequence of TIFF2025096309000031.tif4128 (SEQ ID NO:66, peptide 6 / 6a consensus). [The present invention 1056] The isolated peptide of the present invention 1045, comprising the amino acid sequence of TIFF2025096309000032.tif4128 (SEQ ID NO:135, P6 / 6a min consensus). [The present invention 1057] (i) X at position 2 can be Q, N, E, γ - glutamate, D, iso - D, T, S, A, or G; The X at the 3rd position can be K, Q, N, E, g-glutamate, D, isoD, T, S, A, or G; The X at the 6th position can be K, Q, N, E, g-glutamate, D, isoD, T, S, A, or G; The X at the 9th position can be E, g-glutamate, D, isoD, Q, N, T, S, A, or G; and The X at the 10th position can be A, G, S, T, E, g-glutamate, D, isoD, Q, or N, TIFF2025096309000033.tif5128 (SEQ ID NO:16, P25 consensus), or (ii) The X at the 2nd position can be T, S, A, G, D, isoD, E, g-glutamate, Q, or N; The X at the 3rd position can be G, T, S, A, D, isoD, E, g-glutamate, Q, or N; The X at the 6th position can be Q, N, E, g-glutamate, D, isoD, T, S, A, or G; The X at the 7th position can be K, Q, N, E, g-glutamate, D, isoD, T, S, A, or G; The X at the 10th position can be K, Q, N, E, g-glutamate, D, isoD, T, S, A, or G; The X at the 13th position can be E, g-glutamate, D, isoD, Q, N, T, S, A, or G; The X at the 14th position can be A, G, S, T, E, g-glutamate, D, isoD, Q, or N; and The V at the 16th position is optional, TIFF2025096309000034.tif5128 (SEQ ID NO:17, P25 consensus) An isolated peptide comprising the amino acid sequence of. [Inventive concept 1058] The isolated peptide of inventive concept 1057, which does not consist of SEQ ID NO:179. [Inventive concept 1059] An isolated peptide of the present invention 1057, which contains one or more mutations as compared with the corresponding wild-type amino acid sequence, wherein the polypeptide containing the corresponding wild-type amino acid sequence is SEQ ID NO: 179, and the isolated peptide is more stable than the polypeptide of SEQ ID NO: 179 when dissolved in water or an aqueous solution. [The present invention 1060] An isolated peptide of the present invention 1057, which contains one or more mutations as compared with the corresponding wild-type amino acid sequence, wherein the polypeptide containing the corresponding wild-type amino acid sequence is SEQ ID NO: 179, and the isolated peptide has higher resistance to chemical decomposition than the polypeptide of SEQ ID NO: 179 when dissolved in an aqueous buffer containing a biocide. [The present invention 1061] An isolated peptide of the present invention 1057, which contains the amino acid sequence of TIFF2025096309000035.tif5128 (SEQ ID NO: 16, P25 consensus). [The present invention 1062] An isolated peptide of the present invention 1057, which contains the amino acid sequence of TIFF2025096309000036.tif5128 (SEQ ID NO: 17, P25 consensus). [The present invention 1063] An isolated peptide of the present invention 1057, which contains the amino acid sequence of TIFF2025096309000037.tif5128 (SEQ ID NO: 17, P25 consensus). [The present invention 1064] An isolated peptide of the present invention 1057, which contains one of the amino acid sequences of SEQ ID NOs: 180 to 188. [The present invention 1065] An isolated peptide of the present invention 1057, which has a length of less than 100 amino acids. [The present invention 1066] An isolated peptide of the present invention 1057, which has a peptide length of 20 to 50. [The present invention 1067] (i) The X at position 1 can be any amino acid, but preferably Q, S, E, γ-glutamate, A, T, G, D, iso-D, N, K, or R; The X at position 2 can be any amino acid, but preferably Q, S, E, γ-glutamate, A, T, G, D, iso-D, N, K, or R; The X at position 3 can be any amino acid, but preferably P, Q, S, E, γ-glutamate, A, T, G, D, iso-D, N, K, or R; The X at position 4 can be any amino acid, but preferably I, Q, S, E, γ-glutamate, A, T, G, D, N, iso-D, K, or R; The X at position 5 can be any amino acid, but preferably D, iso-D, S, E, γ-glutamate, A, T, G, N, Q, K, or R; The X at position 6 can be any amino acid, but preferably R, Q, S, E, γ-glutamate, A, T, G, D, iso-D, N, or K; The X at position 7 can be any amino acid, but preferably Q, S, E, γ-glutamate, A, T, G, D, iso-D, N, K, or R; The X at position 8 can be any amino acid, but preferably T, Q, S, E, γ-glutamate, A, G, D, iso-D, N, K, or R; The X at position 9 can be any amino acid, but preferably I, Q, S, E, γ-glutamate, A, T, G, D, iso-D, N, K, or R; The X at position 10 can be any amino acid, but preferably E, γ-glutamate, Q, S, A, T, G, D, iso-D, N, K, or R; The X at position 11 can be any amino acid, but preferably Q, S, E, γ-glutamate, A, T, G, D, iso-D, N, K, or R; The X at position 13 can be any amino acid, but preferably A, S, T, G, D, iso-D, E, γ-glutamate, Q, N, K, or R; The X at position 14 can be any amino acid, but preferably Q, A, S, T, G, D, isoD, E, γ-glutamate, N, K, or R; The X at position 17 can be any amino acid, but preferably A, S, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; The X at position 18 can be any amino acid, but preferably Q, A, S, T, G, D, isoD, E, γ-glutamate, N, K, or R; The X at position 21 can be any amino acid, but preferably K, A, S, T, G, D, isoD, E, γ-glutamate, Q, N, or R; The X at position 22 can be any amino acid, but preferably S, A, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; The X at position 25 can be any amino acid, but preferably S, A, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; The X at position 26 can be any amino acid, but preferably P, S, A, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; and The X at position 27 can be any amino acid, but preferably Q, S, A, T, G, D, isoD, E, γ-glutamate, N, K, or R, TIFF2025096309000038.tif5134 (SEQ ID NO:18, P17 / 18), or (ii) The X at position 2 can be any amino acid, but preferably A, S, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; The X at position 3 can be any amino acid, but preferably Q, A, S, T, G, D, isoD, E, γ-glutamate, N, K, or R; The X at position 6 can be any amino acid, but preferably A, S, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; The X at position 7 can be any amino acid, but preferably Q, A, S, T, G, D, isoD, E, γ-glutamate, N, K, or R; The 10th X can be any amino acid, but preferably K, A, S, T, G, D, isoD, E, γ-glutamate, Q, N, or R; and the 11th X can be any amino acid, but preferably S, A, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R, TIFF2025096309000039.tif5128 (SEQ ID NO:25, P17 / 18min consensus) contains the amino acid sequence of contains one or more mutations compared to the corresponding wild-type amino acid sequence, and the one or more mutations improve the water solubility, stability, or resistance to chemical degradation of the isolated peptide compared to the polypeptide containing the corresponding wild-type amino acid sequence, said isolated peptide. [Inventive Item 1068] The isolated peptide of Inventive Item 1067, which does not consist of SEQ ID NO:162. [Inventive Item 1069] The isolated peptide of Inventive Item 1067, wherein the polypeptide containing the corresponding wild-type amino acid sequence is SEQ ID NO:162, and the isolated peptide is more stable than the polypeptide of SEQ ID NO:162 when dissolved in water or an aqueous solution. [Inventive Item 1070] The isolated peptide of Inventive Item 1067, wherein the polypeptide containing the corresponding wild-type amino acid sequence is SEQ ID NO:162, and the isolated peptide has higher resistance to chemical degradation than the polypeptide of SEQ ID NO:162 when dissolved in an aqueous buffer containing a biocide. [Inventive Item 1071] The isolated peptide of Inventive Item 1067, which contains one of the amino acid sequences of SEQ ID NO:84 - 88, 163 - 167, 228, 229, and 231. [Inventive Item 1072] An isolated peptide of the present invention 1067, comprising the amino acid sequence of TIFF2025096309000040.tif5128 (SEQ ID NO:18, P17 / 18 consensus). [The present invention 1073] An isolated peptide of the present invention 1067, comprising the amino acid sequence of TIFF2025096309000041.tif5128 (SEQ ID NO:25, P17 / 18min consensus). [The present invention 1074] An isolated peptide of the present invention 1067, having less than 100 amino acids in length. [The present invention 1075] An isolated peptide of the present invention 1067, having a peptide length of 20 to 50. [The present invention 1076] Comprising the amino acid sequence of TIFF2025096309000042.tif5128 (SEQ ID NO:26, P19 consensus), where X at position 1 is optional and can be L, I, V, F, or M; X at position 3 is any amino acid, but preferably can be K, A, S, T, G, D, isoD, E, g-glutamate, Q, N, or R; X at position 4 is any amino acid, but preferably can be A, S, T, G, D, isoD, E, g-glutamate, Q, N, K, or R; X at position 7 is any amino acid, but preferably can be K, A, S, T, G, D, isoD, E, g-glutamate, Q, N, or R; X at position 10 is any amino acid, but preferably can be A, S, T, G, D, isoD, E, g-glutamate, Q, N, K, or R; and X at position 11 is any amino acid, but preferably can be R, A, S, T, G, D, isoD, E, g-glutamate, Q, N, or K, an isolated peptide. [The present invention 1077] An isolated peptide of the present invention 1076, which contains one or more mutations as compared with the corresponding wild-type amino acid sequence, the polypeptide containing the corresponding wild-type amino acid sequence is SEQ ID NO:89, and the isolated peptide is more stable than the polypeptide of SEQ ID NO:89 when dissolved in water or an aqueous solution. [The present invention 1078] An isolated peptide of the present invention 1076, which contains one or more mutations as compared with the corresponding wild-type amino acid sequence, the polypeptide containing the corresponding wild-type amino acid sequence is SEQ ID NO:89, and the isolated peptide has higher resistance to chemical decomposition than the polypeptide of SEQ ID NO:89 when dissolved in an aqueous buffer containing a biocide. [The present invention 1079] An isolated peptide of the present invention 1076, which contains one of the amino acid sequences of SEQ ID NO:90-92, 168-173, and 226. [The present invention 1080] An isolated peptide of the present invention 1076, which has a length of less than 100 amino acids. [The present invention 1081] An isolated peptide of the present invention 1076, which has a peptide length of 20-50. [The present invention 1082] An isolated peptide of the present invention 1076, in which the amino acid residue at the 1st position is present. [The present invention 1083] (i) X at the 1st position can be Q, N, D, E, γ-glutamate, isoD, or S; X at the 2nd position can be D, E, γ-glutamate, isoD; X at the 3rd position can be P, D, E, isoD, or γ-glutamate; X at the 4th position can be M, A, S, D, E, isoD, or γ-glutamate; X at the 5th position can be Q, E, or γ-glutamate; X at the 6th position can be A, E, or γ-glutamate; The 8-position X can be M, L, E, Q, D, N, G, A, S, isoD, or g-glutamate; The 9-position X can be Q, N, E, D, G, A, S, isoD, or g-glutamate; The 12-position X can be Q, N, E, D, G, A, S, isoD, or g-glutamate; The 13-position X can be Q, N, E, D, G, A, S, isoD, or g-glutamate; and The 16-position X can be K, Q, N, E, D, R, G, A, or S, TIFF2025096309000043.tif4128 (SEQ ID NO:13, P14d consensus), or (ii) The 2-position X can be M, L, E, Q, D, N, G, A, S, isoD, or g-glutamate; The 3-position X can be Q, N, E, D, G, A, S, isoD, or g-glutamate; The 6-position X can be Q, N, E, D, G, A, S, isoD, or g-glutamate; The 7-position X can be Q, N, E, D, G, A, S, isoD, or g-glutamate; and The 10-position X can be K, Q, N, E, D, R, G, A, or S, TIFF2025096309000044.tif4128 (SEQ ID NO:14, P14d min consensus) An isolated peptide comprising the amino acid sequence of. [Inventive concept 1084] An isolated peptide of inventive concept 1083 that does not consist of SEQ ID NO:174. [Inventive concept 1085] An isolated peptide of inventive concept 1083 that contains one or more mutations compared to the corresponding wild-type amino acid sequence, wherein the polypeptide comprising the corresponding wild-type amino acid sequence is SEQ ID NO:174, and the isolated peptide is more stable than the polypeptide of SEQ ID NO:174 when dissolved in water or an aqueous solution. [Inventive concept 1086] Comprising one or more mutations as compared to the corresponding wild-type amino acid sequence, wherein the polypeptide comprising the corresponding wild-type amino acid sequence is SEQ ID NO: 174, and the isolated peptide has higher resistance to chemical degradation than the polypeptide of SEQ ID NO: 174 when dissolved in an aqueous buffer containing a biocide, the isolated peptide of the present invention 1083. [The present invention 1087] The isolated peptide of the present invention 1083, comprising one of the amino acid sequences of SEQ ID NOs: 175 to 178 and 199. [The present invention 1088] The isolated peptide of the present invention 1083, comprising the amino acid sequence of TIFF2025096309000045.tif4128 (SEQ ID NO: 13, P14d consensus). [The present invention 1089] The isolated peptide of the present invention 1083, comprising the amino acid sequence of TIFF2025096309000046.tif4128 (SEQ ID NO: 14, P14d min consensus). [The present invention 1090] The isolated peptide of the present invention 1083, having a length of less than 100 amino acids. [The present invention 1091] The isolated peptide of the present invention 1083, having a peptide length of 12 to 50. [The present invention 1092] (i) X at position 2 can be Q, N, E, γ-glutamate, D, isoD, T, S, A, or G; X at position 3 can be Q, N, E, γ-glutamate, D, isoD, T, S, A, or G; X at position 6 can be K, Q, N, E, γ-glutamate, D, isoD, T, S, A, or G; X at position 9 can be E, γ-glutamate, D, isoD, Q, N, T, S, A, or G; X at position 10 can be A, G, S, T, E, γ-glutamate, D, isoD, Q, or N; The 13-position X can be Q, N, E, g-glutamate, D, isoD, T, S, A, or G; and The 14-position X can be Q, N, E, g-glutamate, D, isoD, T, S, A, or G, An isolated peptide comprising the amino acid sequence of TIFF2025096309000047.tif5128 (SEQ ID NO:15, P3min consensus). [Inventive concept 1093] An isolated peptide of inventive concept 1092 that does not consist of the 31 amino acid portion from SEQ ID NO:230 and its C-terminus. [Inventive concept 1094] An isolated peptide of inventive concept 1092 that contains one or more mutations compared to the corresponding wild-type amino acid sequence, wherein the polypeptide containing the corresponding wild-type amino acid sequence is SEQ ID NO:230, and the isolated peptide is more stable than the polypeptide of SEQ ID NO:230 when dissolved in water or an aqueous solution. [Inventive concept 1095] An isolated peptide of inventive concept 1092 that contains one or more mutations compared to the corresponding wild-type amino acid sequence, wherein the polypeptide containing the corresponding wild-type amino acid sequence is SEQ ID NO:230, and the isolated peptide has higher resistance to chemical degradation than the polypeptide of SEQ ID NO:230 when dissolved in an aqueous buffer containing a biocide. [Inventive concept 1096] An isolated peptide of inventive concept 1092 that contains one or more mutations compared to the corresponding wild-type amino acid sequence, wherein the polypeptide containing the corresponding wild-type amino acid sequence is SEQ ID NO:230, and the isolated peptide has higher solubility in an aqueous solution than the polypeptide of SEQ ID NO:230. [Inventive concept 1097] An isolated peptide of inventive concept 1092 consisting of SEQ ID NO:204. [Inventive concept 1098] An isolated peptide of the present invention 1092, comprising one amino acid sequence among SEQ ID NOs: 205 to 209. [The present invention 1099] An isolated peptide of the present invention 1092, having a length of less than 100 amino acids. [The present invention 1100] An isolated peptide of the present invention 1092, having a peptide length of 12 to 30. [The present invention 1101] An isolated peptide of any one of the present inventions 1001 to 1100, being at least 90% pure. [The present invention 1102] A fusion polypeptide comprising a second amino acid sequence coupled to the amino acid sequence via a peptide bond, which is an isolated peptide of any one of the present inventions 1001 to 1030, 1033, 1036 to 1040, 1044 to 1051, 1054 to 1096, and 1098 to 1100. [The present invention 1103] An isolated peptide of the present invention 1102, wherein the second amino acid sequence contains a purification tag. [The present invention 1104] An isolated peptide of the present invention 1103, wherein the second amino acid sequence further contains a cleavable linker sequence between the purification tag and the amino acid sequence. [The present invention 1105] An isolated peptide of the present invention 1102, which is a fusion polypeptide comprising the first amino acid sequence of the peptide linked to the second amino acid sequence of the peptide. [The present invention 1106] An isolated peptide of the present invention 1102, wherein the second amino acid sequence contains an N-terminal or C-terminal hydrophilic amino acid sequence. [The present invention 1107] An isolated peptide of the present invention 1106, wherein the hydrophilic amino acid sequence contains a plurality of Glu (E) amino acid residues. [The present invention 1108] A fusion polypeptide comprising a plurality of amino acid sequences linked together continuously, each of the plurality of amino acid sequences comprising any one of the peptides of the present invention from 1001 to 1030, 1033, 1036 to 1040, 1044 to 1051, 1054 to 1096, and 1098 to 1100. [The present invention 1109] The fusion polypeptide of the present invention 1108, wherein the plurality of amino acid sequences are linked together by a cleavable linker sequence. [The present invention 1110] The fusion polypeptide of the present invention 1108, wherein each of the plurality of amino acid sequences comprises a purification tag, an N-terminal or C-terminal hydrophilic amino acid sequence, or both. [The present invention 1111] A composition comprising one or more peptides of any one of the present invention from 1001 to 1107 or a fusion polypeptide of any one of the present invention from 1108 to 1110 and a carrier. [The present invention 1112] The composition of the present invention 1111, which is a clarified cell extract. [The present invention 1113] The composition of the present invention 1111, further comprising an additive selected from the group consisting of fertilizers, herbicides, insecticides, fungicides, nematicides, bactericides, biological inoculants, plant regulators, and mixtures thereof. [The present invention 1114] The composition of the present invention 1113, wherein the insecticide is a neonicotinoid insecticide, an organophosphate insecticide, a pyrethroid insecticide, a macrocyclic lactone insecticide, a carbamate insecticide, a diamide insecticide, an abamectin insecticide, a chitin synthesis inhibitor, or any combination thereof. [The present invention 1115] The composition of the present invention 1113, wherein the fungicide is a strobilurin fungicide, a triazole fungicide, a succinate dehydrogenase fungicide, a phenylamide fungicide, a phenylpyrrole fungicide, a phthalimide fungicide, a dithiocarbamate fungicide, a benzimidazole fungicide, or any combination thereof. [The present invention 1116] The composition of the present invention 1113, wherein the nematicide is a carbamate nematicide. [The present invention 1117] The composition of the present invention 1113, wherein the fungicide is dichlorophen and benzyl alcohol hemi formaldehyde fungicide, isothiazolinone fungicide, or a combination thereof. [The present invention 1118] The composition of the present invention 1113, wherein the biological inoculum is of the genus Bradyrhizobium, Bacillus, Streptomyces, Trichoderma, Pasteuria, or any combination thereof. [The present invention 1119] One or more of the peptides P1, P4-14S, P6a, P14d, P15a, P18, P19, or P25; and clothianidin, a combination of clothianidin and Bacillus firmus, imidacloprid, or a combination of imidacloprid and Bacillus firmus The composition of the present invention 1113 comprising the same. [The present invention 1120] One or more of the peptides P1, P4-14S, P6a, P14d, P15a, P18, P19, or P25, and thiamethoxam; a combination of thiamethoxam, mefenoxam, and fludioxonil; a combination of thiamethoxam, mefenoxam, fludioxonil, and azoxystrobin; a combination of thiamethoxam and abamectin; a combination of thiamethoxam, abamectin, and a Pasteuria nematicide; or a combination of thiamethoxam, mefenoxam, fludioxonil, azoxystrobin, thiabendazole, and abamectin The composition of the present invention 1113 comprising the same. [The present invention 1121] One or more of the peptides P1, P4-14S, P6a, P14d, P15a, P18, P19, or P25; and Biological inoculants including Bradyrhizobium, Bacillus, and combinations thereof The composition of the present invention 1113 comprising the same. [The present invention 1122] The composition of the present invention 1111, wherein the carrier is an aqueous carrier. [The present invention 1123] The composition of the present invention 1122, wherein the aqueous carrier further comprises one or more of a biocide, a protease inhibitor, a nonionic surfactant, or a combination thereof. [The present invention 1124] The composition of the present invention 1111, wherein the carrier is a solid carrier in particulate form. [The present invention 1125] The composition of the present invention 1124, wherein the solid carrier is a dry powder. [The present invention 1126] Applying an effective amount of any of the isolated peptides of the present invention 1001 to 1107, any of the fusion polypeptides of the present invention 1108 to 1110, or any of the compositions of the present invention 1111 to 1125 to a plant or a plant seed or to a location where the plant is growing or is predicted to grow A method for conferring disease resistance to a plant, comprising the above application, wherein the application is effective for conferring disease resistance. [The present invention 1127] The method of the present invention 1126, wherein the disease is a viral disease, a bacterial disease, or a fungal disease. [The present invention 1128] The method of the present invention 1126, wherein the application is carried out using a plant. [The present invention 1129] The method of the present invention 1128, wherein the plant is resistant to at least one herbicide. [The present invention 1130] The method of the present invention 1126, wherein the application is carried out using a plant seed, and the method further comprises planting the seed treated with the peptide or the composition in natural soil or artificial soil, and propagating a plant from the seed planted in the soil. [The present invention 1131] The method of the present invention 1126, wherein the application is carried out at the location where the plant is growing or is predicted to grow. [The present invention 1132] The method of the present invention 1126, wherein the plant is selected from agricultural plants, forestry plants, ornamental plants, and horticultural plants, each in its natural form or genetically modified form. [The present invention 1133] The method of the present invention 1126, wherein the plant is a genetically modified plant. [The present invention 1134] The plant to be treated is selected from the group consisting of alfalfa, apple, apricot, asparagus, avocado, banana, barley, bean, beech (Fagus species), begonia, camphor, canola, carrot, castor bean, cherry, Chinese cinnamon, citrus, cocoa bean, coffee, corn, cotton, cucumber, gourd, eucalyptus, fir, flax, fodder beet, fuchsia, garlic, foxglove, grape, peanut, hemp, hop, Japanese elm, Brassica juncea, jute, lentil, lettuce, linseed, melon, mustard, oak, oats, oil palm, rape, olive, onion, paprika, pea, peach, pear, pelargonium, pepper, petunia, pine (Pinus species), poplar (Populus species), pome fruit, potato, rape, raspberry, rice, rubber tree, rye, sorghum, soybean, spinach, spruce, squash, strawberry, sugar beet, sugarcane, sunflower, tea tree, teak, tobacco, tomato, triticale, turfgrass, watermelon, wheat, and willow (Salix species). [The present invention 1135] Applying an effective amount of any of the isolated peptides of the present invention 1001 to 1107, any of the fusion polypeptides of the present invention 1108 to 1110, or any of the compositions of the present invention 1111 to 1125 to a plant or plant seed or to the location where the plant is growing or is predicted to grow A method for promoting plant growth, comprising the application and being effective for promoting plant growth. [Invention 1136] The method of Invention 1135, wherein the promotion of growth includes an increase in plant vigor, an increase in plant weight, an increase in biomass, an increase in the number of flowers per plant, a higher yield of grains and / or fruits, more tillers or side shoots, larger leaves, an increase in shoot growth, an increase in protein content, an increase in oil content, an increase in starch content, an increase in pigment content, an increase in chlorophyll content, and combinations thereof. [Invention 1137] The method of Invention 1135, wherein the application is carried out using a plant. [Invention 1138] The method of Invention 1137, wherein the plant is resistant to at least one herbicide. [Invention 1139] The method of Invention 1135, wherein the application is carried out using plant seeds, and the method further includes planting the seeds treated with the peptide or the composition in natural soil or artificial soil, and propagating a plant from the seeds planted in the soil. [Invention 1140] The method of Invention 1135, wherein the application is carried out at a location where the plant is growing or is predicted to grow. [Invention 1141] The method of Invention 1135, wherein the plant is selected from agricultural plants, forestry plants, ornamental plants, and horticultural plants in their natural form or genetically modified form, respectively. [Invention 1142] The method of Invention 1135, wherein the plant is a genetically modified plant. [Invention 1143] The method of the present invention 1135, wherein the plant to be treated is selected from the group consisting of alfalfa, apple, apricot, asparagus, avocado, banana, barley, bean, beech (Fagus species), begonia, boxwood, blackberry, blueberry, cabbage, elm, canola, carrot, sesame, cherry, Japanese apricot, citrus, cocoa bean, coffee, corn, cotton, cucumber, gourd, eucalyptus, fir, flax, fodder beet, fuchsia, garlic, foxglove, grape, peanut, hemp, hop, zephyranthes, Brassica juncea (Indian mustard), jute, lentil, lettuce, ramie, melon, mustard, oak, oat, oil palm, rape, olive, onion, paprika, pea, peach, pear, pelargonium, pepper, petunia, pine (Pinus species), poplar (Populus species), pome fruit, potato, Brassica napus, raspberry, rice, rubber tree, rye, sorghum, soybean, spinach, spruce, pumpkin, strawberry, sugar beet, sugarcane, sunflower, tea tree, teak, tobacco, tomato, triticale, turfgrass, watermelon, wheat, and willow (Salix species). [The present invention 1144] Applying an effective amount of any of the isolated peptides of the present invention 1001 to 1107, any of the fusion polypeptides of the present invention 1108 to 1110, or any of the compositions of the present invention 1111 to 1125 to a plant or plant seed or to a location where the plant is growing or is predicted to grow A method for increasing the tolerance of a plant to biological stress, comprising the above application, wherein the application is effective to increase the tolerance of the plant to biological stress factors selected from the group consisting of insects, spiders, nematodes, weeds, and combinations thereof. [The present invention 1145] The method of the present invention 1144, wherein the application is carried out using a plant. [The present invention 1146] The method of the present invention 1145, wherein the plant is resistant to at least one herbicide. [The present invention 1147] Said application is carried out using plant seeds, and said method further comprises planting said seeds treated with said peptide or said composition in natural soil or artificial soil, and propagating a plant from said seeds planted in said soil, the method of the present invention 1144. [The present invention 1148] The method of the present invention 1144, wherein said application is carried out at a location where the plant is growing or is predicted to grow. [The present invention 1149] The method of the present invention 1144, wherein said plants are each selected from agricultural plants, forestry plants, ornamental plants, and horticultural plants in their natural form or genetically modified form. [The present invention 1150] The method of the present invention 1144, wherein said plant is a genetically modified plant. [The present invention 1151] The plant to be treated is selected from the group consisting of alfalfa, apple, apricot, asparagus, avocado, banana, barley, bean, beech (Fagus species), begonia, birch, blackberry, blueberry, cabbage, elm, canola, carrot, castor bean, cherry, Japanese cypress, citrus, cocoa bean, coffee, corn, cotton, cucumber, gourd, eucalyptus, fir, flax, fodder beet, fuchsia, garlic, foxglove, grape, peanut, hemp, hop, zelkova, brassica juncea (Indian mustard), jute, lentil, lettuce, ramie, melon, mustard, oak, oat, oil palm, rape, olive, onion, paprika, pea, peach, pear, pelargonium, pepper, petunia, pine (Pinus species), poplar (Populus species), pome fruit, potato, rape, raspberry, rice, rubber tree, rye, sorghum, soybean, spinach, spruce, pumpkin, strawberry, sugar beet, sugarcane, sunflower, tea tree, teak tree, tobacco, tomato, triticale, turfgrass, watermelon, wheat, and willow (Salix species), the method of the present invention 1144. [The present invention 1152] Applying an effective amount of any of the isolated peptides of the present invention from 1001 to 1107, any of the fusion polypeptides of the present invention from 1108 to 1110, or any of the compositions of the present invention from 1111 to 1125 to a plant or plant seed or to the location where the plant is growing or is predicted to grow A method for increasing the tolerance of a plant to abiotic stress, comprising the above application, wherein the application is effective for increasing the tolerance of the plant to an abiotic stress factor selected from the group consisting of salt stress, water stress, ozone stress, heavy metal stress, low temperature stress, high temperature stress, nutrient stress, and combinations thereof. [The present invention 1153] The method of the present invention 1152, wherein the applying is carried out using a plant. [The present invention 1154] The method of the present invention 1153, wherein the plant is tolerant to at least one herbicide. [The present invention 1155] The method of the present invention 1152, wherein the applying is carried out using a plant seed, and the method further comprises planting the seeds treated with the peptide or the composition in natural soil or artificial soil, and propagating a plant from the seeds planted in the soil. [The present invention 1156] The method of the present invention 1152, wherein the applying is carried out at the location where the plant is growing or is predicted to grow. [The present invention 1157] The method of the present invention 1152, wherein the plant is selected from agricultural plants, forestry plants, ornamental plants, and horticultural plants, each in its natural form or genetically modified form. [The present invention 1158] The method of the present invention 1152, wherein the plant is a genetically modified plant. [The present invention 1159] The method of the present invention 1152, wherein the plant to be treated is selected from the group consisting of alfalfa, apple, apricot, asparagus, avocado, banana, barley, bean, beech (Fagus species), begonia, boxwood, blackberry, blueberry, cabbage, elm, canola, carrot, sesame, cherry, nettle, citrus, cocoa bean, coffee, corn, cotton, cucumber, gourd, eucalyptus, fir, flax, fodder beet, fuchsia, garlic, plantain, grape, peanut, hemp, hop, zephyranthes, brassica juncea (Indian mustard), jute, lentil, lettuce, ramie, melon, mustard, oak, oat, oil palm, rape, olive, onion, paprika, pea, peach, pear, pelargonium, pepper, petunia, pine (Pinus species), poplar (Populus species), pome fruit, potato, rape, raspberry, rice, rubber tree, rye, sorghum, soybean, spinach, spruce, pumpkin, strawberry, sugar beet, sugarcane, sunflower, tea tree, teak, tobacco, tomato, triticale, turfgrass, watermelon, wheat, and willow (Salix species). [The present invention 1160] Applying an effective amount of any of the isolated peptides of the present invention 1001 to 1107, any of the fusion polypeptides of the present invention 1108 to 1110, or any of the compositions of the present invention 1111 to 1125 to a plant or to a location where the plant is growing A method for imparting drought resistance to an excised portion taken from an ornamental plant, comprising the above application, wherein the application is effective for imparting drought resistance to the excised portion taken from the ornamental plant. [The present invention 1161] The method of the present invention 1160, wherein the applying is carried out using an ornamental plant. [The present invention 1162] The method of the present invention 1160, wherein the applying is carried out at a location where the ornamental plant is growing. [The present invention 1163] The method of the present invention 1160, wherein the ornamental plant is a genetically modified ornamental plant. [The present invention 1164] The method of the present invention 1160, wherein the plant to be treated is selected from the group consisting of beech (Fagus species), begonia, maple, ornamental cabbage, fir, fuchsia, garlic, bugbane, oak, ornamental onion, Pelargonium, petunia, pine (Pinus species), poplar (Populus species), sunflower, teak, tobacco, lawn grass, and willow (Salix species). [The present invention 1165] Applying an effective amount of any of the isolated peptides of the present invention from 1001 to 1107, any of the fusion polypeptides of the present invention from 1108 to 1110, or any of the compositions of the present invention from 1111 to 1125 to a plant containing fruits or vegetables or to the location where the plant is growing, or Applying an effective amount of the isolated peptide or the composition to the harvested fruits or vegetables A method for imparting post-harvest disease resistance or post-harvest drying resistance to fruits or vegetables, comprising the above application, wherein the application is effective for imparting post-harvest disease resistance or post-harvest drying resistance to the fruits or vegetables. [The present invention 1166] The method of the present invention 1165, wherein the application is carried out using a plant. [The present invention 1167] The method of the present invention 1166, wherein the plant is resistant to at least one herbicide. [The present invention 1168] The method of the present invention 1165, wherein the application is carried out at the location where the plant is growing. [The present invention 1169] The method of the present invention 1165, wherein the application is carried out using the harvested fruits or vegetables. [The present invention 1170] The method of the present invention 1165, wherein the plant is a genetically modified plant. [The present invention 1171] The method of the present invention 1165, wherein the plant is selected from the group consisting of apple, apricot, asparagus, avocado, banana, blackberry, blueberry, cabbage, carrot, cherry, citrus, corn, cucumber, gourd, fodder beet, garlic, grape, zephyr lily, juncea (western mustard), lettuce, melon, mustard, olive, onion, pea, peach, pear, pepper, pome fruit, potato, rapeseed, raspberry, spinach, pumpkin, strawberry, sugar beet, sugarcane, tea tree, tomato, rye, and watermelon. [The present invention 1172] Applying an effective amount of any of the isolated peptides of the present invention 1001 to 1107, any of the fusion polypeptides of the present invention 1108 to 1110, or any of the compositions of the present invention 1111 to 1125 to a plant containing fruit or vegetables, or to the location where the plant is growing, or Applying an effective amount of the isolated peptide or the composition to the harvested fruit or vegetables A method for extending the shelf life of fruits or vegetables, comprising the above application, wherein the application is effective for extending the shelf life of fruits or vegetables. [The present invention 1173] The method of the present invention 1172, wherein the application is carried out using a plant. [The present invention 1174] The method of the present invention 1172, wherein the plant is resistant to at least one herbicide. [The present invention 1175] The method of the present invention 1172, wherein the application is carried out at the location where the plant is growing. [The present invention 1176] The method of the present invention 1172, wherein the application is carried out using the harvested fruit or vegetables. [The present invention 1177] The method of the present invention 1172, wherein the plant is a genetically modified plant. [The present invention 1178] The method of the present invention 1172, wherein the plant is selected from the group consisting of apple, apricot, asparagus, avocado, banana, blackberry, blueberry, cabbage, carrot, cherry, citrus, corn, cucumber, gourd, fodder beet, garlic, grape, zephyr lily, juncea (Brassica juncea), lettuce, melon, mustard, olive, onion, pea, peach, pear, pepper, pome fruit, potato, Brassica napus, raspberry, spinach, pumpkin, strawberry, sugar beet, sugarcane, tea tree, tomato, rye, and watermelon. [The present invention 1179] Applying an effective amount of any of the isolated peptides of the present invention 1001 to 1107, any of the fusion polypeptides of the present invention 1108 to 1110, or any of the compositions of the present invention 1111 to 1125 to a plant or plant seed or to a location where the plant is growing or is predicted to grow A method of modulating plant biochemical signal transduction, comprising the above application and being effective for modulating plant biochemical signal transduction. [The present invention 1180] The method of the present invention 1179, wherein the plant biochemical signal transduction is selected from the group consisting of induction of nitric oxide production, peroxide production, or secondary metabolite induction; agonist modulation of the ethylene signal transduction pathway and induction of ethylene response gene expression; agonist modulation of the salicylic acid signal transduction pathway and induction of salicylic acid response gene expression; agonist modulation of the abscisic acid pathway and induction of abscisic acid response gene expression; agonist modulation of the gibberellin signal transduction pathway and induction of gibberellin response gene expression; antagonist modulation of jasmonic acid signal transduction and inhibition of jasmonic acid response gene expression; induction of protease inhibitor expression; induction of reactive oxygen species production in plant tissues; induction of production of immune-related peptides and antimicrobial peptides; and induction of expansin gene expression and production. [The present invention 1181] A DNA construct comprising a first nucleic acid molecule encoding an isolated peptide of any one of polypeptides 1001 to 1107 of the present invention or a fusion polypeptide of any one of polypeptides 1108 to 1110 of the present invention, and a nucleic acid molecule effective as a promoter operably coupled to the first nucleic acid molecule. [Inventive concept 1182] A recombinant expression vector comprising the DNA construct of inventive concept 1181. [Inventive concept 1183] A recombinant host cell comprising the DNA construct of inventive concept 1181. [Inventive concept 1184] The recombinant host cell of inventive concept 1183, which is a plant protoplast. [Inventive concept 1185] The recombinant host cell of inventive concept 1183, which is a bacterium. [Inventive concept 1186] A transformed plant comprising the recombinant host cell of inventive concept 1183. [Inventive concept 1187] A transformed plant seed comprising the recombinant host cell of inventive concept 1183. [Inventive concept 1188] A transformed plant comprising the DNA construct of inventive concept 1181. [Inventive concept 1189] A transformed plant seed comprising the DNA construct of inventive concept 1181. [Inventive concept 1190] Providing a transformed plant transformed with the DNA construct of inventive concept 1181; and Growing the plant under conditions effective for the DNA construct to express the peptide or the fusion polypeptide to confer disease resistance. A method for conferring disease resistance to a plant, comprising the above steps. [Inventive concept 1191] Providing a transformed plant transformed with the DNA construct of inventive concept 1181; and Growing the plant under conditions effective for the DNA construct to express the peptide or the fusion polypeptide and enable enhancement of plant growth A method for enhancing plant growth, comprising: [Inventive concept 1192] Providing a transformed plant transformed with the DNA construct of Inventive concept 1181; and Growing the plant under conditions effective for the DNA construct to express the peptide or the fusion polypeptide and enable imparting of disease resistance A method for imparting disease resistance to a plant, comprising: [Inventive concept 1193] Providing a transformed plant transformed with the DNA construct of Inventive concept 1181; and Growing the plant under conditions effective for the DNA construct to express the peptide or the fusion polypeptide and enable increasing the tolerance of the plant to a biological stress factor selected from the group consisting of insects, spiders, nematodes, weeds, and combinations thereof A method for increasing the tolerance of a plant to biological stress, comprising: [Inventive concept 1194] Providing a transformed plant transformed with the DNA construct of Inventive concept 1181; and Growing the plant under conditions effective for the DNA construct to express the peptide or the fusion polypeptide and enable increasing the tolerance of the plant to an abiotic stress factor selected from the group consisting of salt stress, drought stress, ozone stress, heavy metal stress, low temperature stress, and combinations thereof A method for increasing the tolerance of a plant to abiotic stress, comprising: [Inventive concept 1195] Providing a transformed ornamental plant transformed with the DNA construct of Inventive concept 1181; and Growing the plant under conditions effective to allow the DNA construct to express the peptide or the fusion polypeptide and impart drought resistance to an excised portion taken from the transformed ornamental plant A method for imparting drought resistance to an excised portion taken from an ornamental plant, the method comprising: [Inventive Step 1196] Providing a transformed plant transformed with the DNA construct of Inventive Step 1181; and Growing the plant under conditions effective to allow the DNA construct to express the peptide or the fusion polypeptide and impart post-harvest disease resistance or post-harvest drought resistance to fruits or vegetables harvested from the transformed plant A method for imparting post-harvest disease resistance or post-harvest drought resistance to fruits or vegetables, the method comprising: [Inventive Step 1197] Providing a transformed plant transformed with the DNA construct of Inventive Step 1181; and Growing the plant under conditions effective to allow the DNA construct to express the peptide or the fusion polypeptide and extend the shelf life of fruits or vegetables harvested from the transformed plant A method for extending the shelf life of fruits or vegetables, the method comprising: [Inventive Step 1198] Providing a transformed plant seed transformed with the DNA construct of Inventive Step 1181; Planting the transformed plant seed in soil; and Propagating a transformed plant from the transformed plant seed under conditions effective to allow the DNA construct to express the peptide or the fusion polypeptide and impart disease resistance A method for imparting disease resistance to a plant, the method comprising: [Inventive Step 1199] Providing a transformed plant seed transformed with the DNA construct of Inventive Step 1181; Planting the transformed plant seeds in soil; and Propagating a transformed plant from the transformed plant seeds such that the DNA construct enables expression of the peptide or the fusion polypeptide to enhance plant growth A method for enhancing plant growth, comprising: [Inventive concept 1200] Providing transformed plant seeds transformed with the DNA construct of Inventive concept 1181; Planting the transformed plant seeds in soil; and Propagating a transformed plant from the transformed plant seeds such that the DNA construct enables expression of the peptide or the fusion polypeptide to confer disease resistance A method for conferring disease resistance to a plant, comprising: [Inventive concept 1201] Providing transformed plant seeds transformed with the DNA construct of Inventive concept 1181; Planting the transformed plant seeds in soil; and Propagating the transformed plant from the transformed plant seeds such that the DNA construct enables expression of the peptide or the fusion polypeptide to increase the tolerance of the transformed plant to biological stress factors selected from the group consisting of insects, spiders, nematodes, weeds, and combinations thereof A method for increasing the tolerance of a plant to biological stress, comprising: [Inventive concept 1202] Providing transformed plant seeds transformed with the DNA construct of Inventive concept 1181; Planting the transformed plant seeds in soil; and Propagating the transformed plant from the transformed plant seeds so that the DNA construct expresses the peptide or the fusion polypeptide and enables the transformed plant to have increased tolerance to abiotic stress factors selected from the group consisting of salt stress, drought stress, ozone stress, heavy metal stress, and low temperature stress, and combinations thereof A method for increasing the tolerance of a plant to abiotic stress, comprising: [Inventive concept 1203] Providing transformed ornamental plant seeds transformed with the DNA construct of Inventive concept 1181; Planting the transformed ornamental plant seeds in soil; and Propagating the transformed ornamental plant from the transformed ornamental plant seeds so that the DNA construct expresses the peptide or the fusion polypeptide and enables the excised portion taken from the transformed ornamental plant to have drought resistance A method for imparting drought resistance to an excised portion taken from an ornamental plant, comprising: [Inventive concept 1204] Providing transformed plant seeds transformed with the DNA construct of Inventive concept 1181; Planting the transformed plant seeds in soil; and Propagating the transformed plant from the transformed plant seeds so that the DNA construct expresses the peptide or the fusion polypeptide and enables the fruits or vegetables harvested from the transformed plant to have post-harvest disease resistance or post-harvest drought resistance A method for imparting post-harvest disease resistance or post-harvest drought resistance to fruits or vegetables, comprising: [Inventive concept 1205] Providing transformed plant seeds transformed with the DNA construct of Inventive concept 1181; Planting the transformed plant seeds in soil; and Propagating the transformed plant from the transformed plant seeds in order to enable the DNA construct to express the peptide or the fusion polypeptide and extend the shelf life of fruits or vegetables harvested from the transformed plant A method for extending the shelf life of fruits or vegetables, comprising: [Invention 1206] Providing a transformed plant seed transformed with the DNA construct of Invention 1181; Planting the transformed plant seed in soil; and Propagating a transformed plant from the transformed plant seed in order to enable the DNA construct to express the peptide or the fusion polypeptide and modulate plant biochemical signal transduction A method for modulating plant biochemical signal transduction, comprising: [Invention 1207] The method of Invention 1206, wherein the plant biochemical signal transduction is selected from the group consisting of induction of nitric oxide production, peroxide production, or secondary metabolite induction; agonist modulation of the ethylene signal transduction pathway and induction of ethylene response gene expression; agonist modulation of the salicylic acid signal transduction pathway and induction of salicylic acid response gene expression; agonist modulation of the abscisic acid pathway and induction of abscisic acid response gene expression; agonist modulation of the gibberellin signal transduction pathway and induction of gibberellin response gene expression; antagonist modulation of jasmonic acid signal transduction and inhibition of jasmonic acid response gene expression; induction of protease inhibitor expression; induction of reactive oxygen species production in plant tissues; induction of production of immune-related peptides and antimicrobial peptides; and induction of expansin gene expression and production. BRIEF DESCRIPTION OF THE DRAWINGS

[0040]

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Mode for Carrying Out the Invention

[0041] One aspect of the present invention relates to a novel peptide having the ability to induce a hypersensitive response in plants and to promote an active plant response that results in one or more of the following traits: disease resistance, growth promotion, tolerance and resistance to biological stressors, tolerance to abiotic stress, desiccation resistance in excised parts taken from ornamental plants, post-harvest disease resistance or post-harvest desiccation resistance in fruits or vegetables harvested from plants, and / or improvement of the fruit or vegetable ripening lifespan for fruits or vegetables harvested from plants.

[0042] As used herein, naturally occurring amino acids are identified by their conventional three-letter and / or one-letter abbreviations, which correspond to the common names of the amino acids according to the following list: alanine (Ala, A), arginine (Arg, R), asparagine (Asn, N), aspartic acid (Asp, D), cysteine (Cys, C), glutamic acid (Glu, E), glutamine (Gln, Q), glycine (Gly, G), histidine (His, H), isoleucine (Ile, I), leucine (Leu, L), lysine (Lys, K), methionine (Met, M), phenylalanine (Phe, F), proline (Pro, P), serine (Ser, S), threonine (Thr, T), tryptophan (Trp, W), tyrosine (Tyr, Y), and valine (Val, V). These abbreviations are recognized in the peptide art and are recommended in biochemical nomenclature by the IUPAC-IUB Commission. Naturally occurring variant forms of amino acids include, but are not limited to, gamma-glutamate (g-Glu) and isoaspartate (iso-Asp or isoD).

[0043] The term "amino acid" further includes analogs, derivatives, and homologs of any specific amino acid mentioned herein, as well as amino acid derivatives that are C-terminally or N-terminally protected (e.g., modified with N-terminal, C-terminal, or side-chain protecting groups including, but not limited to, acetylation, formylation, methylation, amidation, esterification, pegylation, and addition of lipids). Non-naturally occurring amino acids are well known and can be introduced into the peptides of the present invention using solid-phase synthesis as described below. Further, the term "amino acid" includes both D- and L-amino acids. Thus, in this specification, an amino acid that is identified by its name, three-letter symbol, or one-letter symbol and is not specifically identified as having either a D or L configuration is understood to assume either a D or L configuration. In one embodiment, the peptide comprises all L-amino acids.

[0044] In certain embodiments, the peptide is identified as "consisting of" the recited sequence, in which case the peptide contains only the recited amino acid sequence without any foreign amino acids at its N-terminus or C-terminus. Multiple peptide sequences are included within peptides that consist of such recited sequences, provided that the recited sequence is in the form of a consensus sequence in which one or more of the indicated X or Xaa residues can be any of one or more amino acids.

[0045] In certain other embodiments, the peptide is identified as "consisting essentially of" the recited sequence, in which case the peptide comprises the recited amino acid sequence optionally including one or more foreign amino acids at its N-terminus and / or C-terminus, where the foreign amino acids do not significantly alter one or more of the following properties: (i) the ability of the peptide to induce a hypersensitive response in plants, (ii) the solubility of the peptide in water or an aqueous solution, (iii) the stability of the peptide dissolved in water or an aqueous solution at 50°C over a period of time (e.g., 3 weeks), and (iv) the resistance of the peptide to chemical degradation in the presence of a biocide (e.g., Proxel® GXL) in an aqueous buffer at 50°C over a period of time (e.g., 3 weeks).

[0046] Briefly, the stability and resistance of a peptide to chemical degradation can be evaluated as follows using a peptide sample having an initial purity of at least about 80%, at least about 82%, at least about 84%, at least about 86%, at least about 88%, at least about 90%, at least about 92%, at least about 94%, at least about 96%, or at least about 98%. For water stability, the peptide is dissolved directly in deionized water. For the chemical degradation test, the peptide is dissolved in an aqueous solution containing 50 mM pH buffer and 0.25% Proxel GXL. Exemplary pH buffers include, but are not limited to: (i) citrate pH 5.6; (ii) MES pH 6.2; (iii) MOPS pH 6.5; (iv) imidazole pH 7.0; (v) citrate pH 7.2; (vi) EDDS, pH 7.3; (vii) EDTA pH 8.0; (viii) sodium phosphate pH 8.0; or (ix) TES pH 8.0. The peptide is first dissolved in an aqueous solution at a concentration of 0.5 mg / ml. The sample is incubated at 50 °C to enable acceleration of degradation. An aliquot of the initial peptide sample is removed, diluted 10-fold with water, and analyzed by reverse-phase HPLC. Briefly, 20 μl of the sample is injected into the solvent flow of an HPLC instrument and analyzed on a C18 HPLC column (YMC ProPack C18, YMC, Japan, or C18 Stablebond, Agilent Technologies, USA) using either a triethylamine phosphate gradient in water / acetonitrile or a gradient of 0.1% TFA in water / 0.1% TFA in acetonitrile to separate different peptide species. Elution of the peptide is monitored by UV absorbance at 218 nm and quantified based on the area under the peak. The area under the peak in the initial peptide sample is treated as a standard for relative quantification in subsequent runs. At regular intervals (e.g., 1, 3, 7, 10, 14, 17, and 21 days), each peptide sample is examined and analyzed by HPLC as described above. If necessary, this protocol can be extended for several weeks to observe degradation (i.e., whether the peptide exhibits a high degree of chemical stability).Quantify the subsequent peptide eluate as a percentage relative to the original (day 0) HPLC results.

[0047] Peptides that are at least partially soluble in water or aqueous solutions exhibit a solubility of greater than 0.1 mg / ml, preferably at least about 1.0 mg / ml, at least about 2.0 mg / ml, at least about 3.0 mg / ml, or at least about 4.0 mg / ml. In certain embodiments, the peptide exhibits high solubility in water or aqueous solutions with a solubility of at least about 5.0 mg / ml, at least about 10.0 mg / ml, at least about 15.0 mg / ml, or at least about 20 mg / ml.

[0048] Peptides that are stable in water or aqueous solutions exhibit at least about 66%, at least about 68%, at least about 70%, at least about 72%, at least about 74%, at least about 76%, at least about 78%, at least about 80%, at least about 82%, at least about 84%, at least about 86%, at least about 88%, or at least about 90% of the original peptide concentration over a specified period of incubation at 50°C. In certain embodiments, the specified period is 3 days, 7 days, 14 days, 21 days, 28 days, 1 month, 2 months, or 3 months.

[0049] Peptides that are resistant to chemical degradation exhibit at least about 66%, at least about 68%, at least about 70%, at least about 72%, at least about 74%, at least about 76%, at least about 78%, at least about 80%, at least about 82%, at least about 84%, at least about 86%, at least about 88%, or at least about 90% of the original peptide concentration over a specified period of incubation at 50°C. In certain embodiments, the specified period is 3 days, 7 days, 14 days, 21 days, 28 days, 1 month, 2 months, or 3 months.

[0050] The property of a peptide to induce or not induce a hypersensitive reaction when infiltrated or applied to plant tissue can be measured by applying the peptide in dry powder form or solution form to the plant, particularly, but not exclusively, to the plant leaves. Application rates include 1 - 500 μg / ml for liquid solutions and 0.0001 - 0.5% (w / w) for powder application. Exemplary applications of the peptide in solution form are described in the attached examples. A plant is judged as HR positive ("HR+") if, within 48 hours, it shows widespread macroscopic cell death visible to the naked eye, accompanied by withering and browning of the affected tissue. A plant is judged as HR negative ("HR-") if it shows no distinguishable withering or tissue death observable by the naked eye.

[0051] In certain embodiments, a significant modification of one or more properties is intended to mean that there is a change of less than 20%, less than 15%, less than 10%, or less than 5% in the described properties when a peptide having one or more foreign amino acids is compared to a peptide that does not contain the one or more foreign amino acids and is otherwise identical. In certain embodiments, the number of foreign amino acids at the N-terminus or C-terminus is at most 20 amino acids at one or both ends, at most 15 amino acids at one or both ends, at most 10 amino acids at one or both ends, at most 7 amino acids at one or both ends, at most 5 amino acids at one or both ends, or at most 3 amino acids at one or both ends. Further, as long as the described sequence is in the form of a consensus sequence where one or more of the indicated X or Xaa residues can be any of one or more amino acids, multiple peptide sequences are encompassed by peptides that consist essentially of such described sequences, regardless of additional modifications of such sequences resulting from the presence of foreign amino acids at their N-terminus and / or C-terminus.

[0052] In various embodiments of the present invention, the disclosed peptides may, for example, include a hydrophilic amino acid sequence either at the N-terminus or the C-terminus of a specified peptide sequence. The hydrophilic amino acid sequence is at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 amino acids in length and includes amino acid residues that contribute to the hydrophilicity of the amino acid sequence adjacent to the amino acid sequence of the specified peptide (i.e., the peptide that induces an active plant response). Various methods are used in the art to calculate the relative hydrophobicity / hydrophilicity of amino acid residues and proteins (Kyte et al., “A Simple Method for Displaying the Hydropathic Character of a Protein,” J. Mol. Biol. 157: 105-32 (1982); Eisenberg D, “Three-dimensional Structure of Membrane and Surface Proteins,” Ann. Rev. Biochem. 53: 595-623 (1984); Rose et al., “Hydrogen Bonding, Hydrophobicity, Packing, and Protein Folding,” Annu. Rev. Biomol. Struct. 22: 381-415 (1993); Kauzmann, “Some Factors in the Interpretation of Protein Denaturation,” Adv. Protein Chem. 14: 1-63 (1959), all of which are incorporated herein by reference in their entirety). Any one of these hydrophobic scales can be used for the purposes of the present invention; however, the Kyte-Doolittle hydrophobic scale is perhaps the most widely referenced scale. These hydrophobic scales provide a ranking list for the relative hydrophobicity of amino acid residues.For example, amino acids that contribute to hydrophilicity include Arg (R), Lys (K), Asp (D), Glu (E), Gln (Q), Asn (N), and His (H), and to a lesser extent, Ser (S), Thr (T), Gly (G), Pro (P), Tyr (Y), and Trp (W). For example, polyglutamate sequences can be used to enhance the solubility of proteins and other drug molecules (Lilie et al, Biological Chemistry 394(8):995 - 1004(2013); Li et al., Cancer Research 58: 2404 - 2409(1998), each of which is incorporated herein by reference in its entirety).

[0053] The "hydropathy index" of a protein or amino acid sequence is a numerical value that represents its average hydrophilicity or hydrophobicity. A negative hydropathy index defines the hydrophilicity of the amino acid sequence in question. Since the hydropathy index is directly proportional to the hydrophilicity of the amino acid sequence in question; the more negative the index, the higher its hydrophilicity. In certain embodiments, the additional hydrophilic amino acid sequences described above have a hydropathy index of less than 0, -0.4, -0.9, -1.3, -1.6, -3.5, -3.9, or -4.5. In certain embodiments, the resulting whole peptide will have a hydropathy index of less than 0.3, 0.2, 0.1, or 0.0, preferably less than -0.1, -0.2, -0.3, -0.4, more preferably less than -0.5, -0.6, -0.7, -0.8, -0.9, or -1.0.

[0054] In the peptide of the present invention, as a whole peptide or in any of the added hydrophilic amino acid sequences, the amino acids contributing to the hydrophilic hydropathy index include Arg (R), Lys (K), Asp (D), Glu (E), Gln (Q), Asn (N), His (H), Ser (S), Thr (T), Gly (G), Pro (P), Tyr (Y), and Trp (W). Among these, Asp (D), Glu (E), Gln (Q), Asn (N), or their variants are preferred. Exemplary variants include γ-glutamate for Glu and isoaspartic acid (or iso-D) for Asp.

[0055] As used herein, in this and other aspects of the present invention, the term "hydrophobic amino acid" is intended to refer to an amino acid that confers hydrophobicity to the hydropathy index of the designated amino acid sequence. Amino acids contributing to the hydrophobic hydropathy index include, as a whole peptide or in any of its specific amino acid sequences, Ile (I), Val (V), Leu (L), Phe (F), Cys (C), Met (M), and Ala (A). In certain embodiments, the term "hydrophobic amino acid" can refer to any one of Ile (I), Val (V), Leu (L), Phe (F), Cys (C), Met (M), and Ala (A); alternatively, in another way, it can refer to any one of Ile (I), Val (V), Leu (L), Phe (F), and Ala (A). In certain other embodiments, the term "hydrophobic amino acid" can refer to one of Ile (I), Val (V), Leu (L), and Phe (F).

[0056] As used herein, the term "non-hydrophobic amino acid" is intended to mean an amino acid that is hydrophilic (or not hydrophobic) on one of the hydrophobic scales identified above. This term generally refers to an amino acid that contributes to the hydrophilic hydropathy index, as a whole peptide or in any of the added hydrophilic amino acid sequences.

[0057] In one aspect of the present invention, the peptide is It contains the amino acid sequence of TIFF2025096309000048.tif5163, where the peptide does not contain cysteine and methionine; each X at positions 2 and 6 is optional and, if present, is any amino acid containing any naturally occurring amino acid; and each X at positions 3, 7, 10, and 11 is any amino acid containing any naturally occurring amino acid.

[0058] In a related aspect of the present invention, the peptide contains the amino acid sequence of SEQ ID NO:93 (shown above), and the peptide does not contain cysteine and methionine; each X at positions 2, 6, and 10 is optional and, if present, is any amino acid containing any naturally occurring amino acid; and each X at positions 3, 7, and 11 is any amino acid containing any naturally occurring amino acid.

[0059] According to one embodiment, one or more of the Xs at positions 2, 6, and 10 are absent (i.e., the gap between the first hydrophobic amino acid and the first hydrophobic amino acid doublet is reduced from 2 to 1 amino acid residue, and / or the gap between the first and second hydrophobic amino acid doublets is reduced from 2 to 1 amino acid residue, and / or the gap between the second and third hydrophobic amino acid doublets is reduced from 2 to 1 amino acid residue). In this embodiment, it is contemplated that these peptides exclude only one of the amino acids at positions 2, 6, and 10.

[0060] In an alternative embodiment, Xs are present at both positions 2 and 6 (i.e., the gaps between the hydrophobic amino acids are maintained at 2 amino acid residues at both positions).

[0061] In another embodiment, an X is present at each of positions 2, 6, and 10 (i.e., the gaps between the hydrophobic amino acids are maintained at 2 amino acid residues at each position).

[0062] In certain embodiments, SEQ ID NO:93 may further include additional amino acid residues between hydrophobic doublets (two of L / I / V / F / A as shown), and the additional amino acids can be any amino acid. In these embodiments, the gap before the first hydrophobic doublet is 3 amino acids, or the gap between the first and second hydrophobic doublets is 3 amino acids, or the gap between the second and third hydrophobic doublets is 3 amino acids, or a combination thereof.

[0063] The peptide length in this embodiment is less than 100 amino acids, or alternatively less than 90 amino acids, less than 80 amino acids, less than 70 amino acids, less than 60 amino acids, or less than about 50 amino acids. In certain embodiments, the peptide length is from 13 to about 50 amino acids in length.

[0064] In the above embodiments, when each of the X's at positions 2, 3, 6, 7, 10, and 11 of SEQ ID NO:93 (if present) can be any amino acid, in certain embodiments, these residues are essentially hydrophilic. As noted above, these hydrophilic amino acids include Arg (R), Lys (K), Asp (D), Glu (E), Gln (Q), Asn (N), His (H), Ser (S), Thr (T), Gly (G), Pro (P), Tyr (Y), and Trp (W). Of these, Asp (D), Glu (E), Gln (Q), Asn (N), or variants thereof are preferred. Exemplary variants include g-glutamate for Glu and isoaspartic acid (or iso-D) for Asp.

[0065] In this embodiment, the isolated peptide is stable when dissolved in water; as noted above, it is resistant to chemical degradation under aqueous conditions in the presence of a pH buffer and a biocide; and / or has a solubility in an aqueous solution of at least about 1.0 mg / ml.

[0066] Another aspect of the invention is Regarding an isolated peptide having the amino acid sequence of TIFF2025096309000049.tif4128 (SEQ ID NO:1, P1 / P4 consensus), where X at position 1 is optional and can be S, N, D, isoD, G, A, or S; X at position 2 is optional and can be Q, E, g-glutamate, G, A, or S; X at position 8 is Q, E, g-glutamate, G, A, or S; X at position 9 is L, I, F, or V; X at position 10 is optional and can be D or isoD; X at position 11 is Q, E, g-glutamate, G, A, or S; X at position 12 is M, L, I, or F; X at position 13 is M, L, or I; X at position 14 is optional and can be any hydrophilic amino acid, preferably C, S, T, A, D, isoD, K, or Q; X at position 15 is Q, E, g-glutamate, G, A, S, K, or I; X at position 16 is M, L, I, V, or F; X at position 17 is M, L, I, A, or V; X at position 18 is Q, E, g-glutamate, G, A, S, M, T, or K; X at position 19 is A, D, isoD, S, V, T, K, R, E, g-glutamate, H, or G; X at position 20 is M, L, or I; X at position 21 is M, L, I, V, S, or F; X at position 22 is Q, E, g-glutamate, G, A, S; X at position 23 is P, Q, E, g-glutamate, G, A, or S, and The isolated peptide contains one or more mutations as compared to the corresponding wild-type amino acid sequence. In certain embodiments, the one or more mutations improve the water solubility, stability, or resistance to chemical degradation of the isolated peptide as compared to a polypeptide containing the corresponding wild-type amino acid sequence.

[0067] In certain embodiments, these peptides according to the second aspect of the invention also have the structural features defining the peptide of SEQ ID NO:93, in which case there are no methionine and cysteine residues.

[0068] In this embodiment, for the purpose of comparing the properties of the peptides of the invention, the corresponding wild-type amino acid sequence is a polypeptide comprising the amino acid sequence of TIFF2025096309000050.tif12128 (amino acids 1 to 21 of SEQ ID NO:5, P4), or a peptide consisting thereof. P1 (SEQ ID NO:4) is derived from the full-length protein of Xanthomonas harpin HpaG (Kim et al., “Mutational Analysis of Xanthomonas Harpin HpaG Identifies a Key Functional Region. That Elicits the Hypersensitive Response in Nonhost Plants,” J. Bacteriol. 186(18):6239-6247 (2004), which is incorporated herein by reference in its entirety). P4 (SEQ ID NO:5) is derived from the full-length harpin of Xanthomonas oryzae pv. oryzae (Ji et al., “Two Coiled-Coil Regions of Xanthomonas oryzae pv. Oryzae Harpin Differ in Oligomerization and Hypersensitive Response Induction,” Amino Acids 40:381-392 (2011), which is incorporated herein by reference in its entirety).

[0069] In this embodiment, the isolated peptide is stable when dissolved in water; as described above, it is resistant to chemical degradation under aqueous conditions in the presence of a pH buffer and a biocide; and / or has a solubility in an aqueous solution of at least about 1.0 mg / ml.

[0070] The length of the peptide according to this second aspect is preferably less than about 100 amino acids, or alternatively less than 90 amino acids, less than 80 amino acids, less than 70 amino acids, less than 60 amino acids, or less than about 50 amino acids. In certain embodiments, the peptide length is from 23 to about 50 amino acids in length.

[0071] One exemplary family of peptides according to the second aspect of the invention is having the amino acid sequence of TIFF2025096309000051.tif4128 (SEQ ID NO:2, P4 consensus), where X at position 2 is Q, E, γ-glutamate, G, A, or S; X at position 8 is Q, E, γ-glutamate, G, A, or S; X at position 9 is L, A, D, isoD, I, V, or F; X at position 11 is Q, E, γ-glutamate, G, A, or S; X at position 12 is L, D, isoD, I, or F; X at position 13 is L, I, V, or F; X at position 14 is any hydrophilic amino acid, preferably C, S, or T, S or T, or S only; X at position 15 is Q, E, γ-glutamate, G, A, S, K, or I; X at position 16 is L, A, I, V, M, or F; X at position 17 is I, S, or F; X at position 18 is Q, E, γ-glutamate, G, A, or S; X at position 20 is L, I, V, or F; The 21st X is L or F; and the 22nd X is Q, E, g-glutamate, G, A, or S.

[0072] In certain embodiments, these peptides according to SEQ ID NO:2 also have the structural features that define the peptide of SEQ ID NO:93, in which case there are no methionine and cysteine residues. Thus, in these embodiments, the 14th X is S or T, preferably S.

[0073] Exemplary peptides that share a consensus structure with SEQ ID NO:2 or are derived from SEQ ID NO:2 and have the consensus structure of SEQ ID NO:93 are identified in Table 1 below.

[0074] (Table 1) Peptide variants of peptide P4 (SEQ ID NO:5) TIFF2025096309000052.tif149134Continuation of Table 1 TIFF2025096309000053.tif222154The selected peptides in Table 1 TIFF2025096309000054.tif4148include solubility tags indicated in italics. Peptides that include the sequences shown in Table 1 but do not include these specific solubility tags (or have different solubility tags) are also contemplated herein.

[0075] As described above, the peptide P4 (SEQ ID NO:5) is derived from the harpin of Xanthomonas oryzae pv. oryzae (Ji et al., “Two Coiled-Coil Regions of Xanthomonas oryzae pv. Oryzae Harpin Differ in Oligomerization and Hypersensitive Response Induction,” Amino Acids 40:381-392 (2011), which is incorporated herein by reference in its entirety). Ji et al. disclose a fragment of this harpin having the amino acid sequence TIFF2025096309000055.tif5128 (i.e., amino acids 1-21 of SEQ ID NO:5). In certain embodiments, the isolated peptide comprising the amino acid sequence of SEQ ID NO:5 is a peptide having an overall length of less than about 100 amino acids (i.e., from amino acid 23 to up to about 100 amino acids in length). In certain other embodiments, the isolated peptide consists essentially of SEQ ID NO:5, while in other embodiments, the isolated peptide consists of SEQ ID NO:5.

[0076] Another exemplary family of peptides according to a second aspect of the invention is TIFF2025096309000056.tif4128 having the amino acid sequence of (SEQ ID NO:3, P1 consensus), where X at position 1 is N, D, isoD, G, A, or S; X at position 2 is Q, E, g-glutamate, G, A, or S; X at position 8 is Q, E, g-glutamate, G, A, or S; X at position 11 is Q, E, g-glutamate, G, A, or S; X at position 15 is Q, E, g-glutamate, G, A, or S; X at position 18 is M, T, K, E, g-glutamate, G, A, or S; X at position 22 is Q, E, g-glutamate, G, A, or S; and The 23-position X is Q, E, g-glutamate, G, A, or S.

[0077] In certain embodiments, these peptides according to SEQ ID NO:3 also have the structural features that define the peptide of SEQ ID NO:93, in which case there are no methionine and cysteine residues. Thus, in these embodiments, the 18-position X is T, K, E, g-glutamate, G, A, or S.

[0078] In certain embodiments, for a peptide sharing the structure of SEQ ID NO:3, at least one of the residues at positions 2, 8, 11, 15, 22, and 23 of SEQ ID NO:3 is other than Gln (Q), i.e., E, g-glutamate, G, A, or S. In certain embodiments, two or more of the residues at positions 2, 8, 11, 15, 22, and 23 of SEQ ID NO:3 are other than Gln (Q), including the case where 3, 4, 5, or all 6 of these residues are other than Gln (Q).

[0079] Exemplary peptides that share a consensus structure with SEQ ID NO:3 or are derived from SEQ ID NO:3 and have the consensus structure of SEQ ID NO:93 are identified in Table 2 below.

[0080] (Table 2) Peptide variants of peptide P1 (SEQ ID NO:4) The selected peptides in Table 2, TIFF2025096309000057.tif82140, contain a solubility tag indicated in italics that includes SEEEEE. Peptides that include the sequences shown in Table 2 but do not contain this particular solubility tag (or have a different solubility tag) are also contemplated herein.

[0081] As described above, the peptide of SEQ ID NO:4 is derived from the harpin of Xanthomonas oryzae pv. oryzae (Ji et al., “Two Coiled-Coil Regions of Xanthomonas oryzae pv. Oryzae Harpin Differ in Oligomerization and Hypersensitive Response Induction,” Amino Acids 40:381-392 (2011), which is incorporated herein by reference in its entirety).

[0082] Another aspect of the present invention is (i) X at position 3 is N, D, or isoD; X at position 6 is Q, E, γ-glutamate, G, A, or S; X at position 8 is N, D, or isoD; X at position 15 is optional and can be any amino acid; X at position 18 is M, E, γ-glutamate, G, A, S, T, or K; and X at position 22 is optional and can be Q, E, γ-glutamate, G, A, or S, TIFF2025096309000058.tif4128 (SEQ ID NO:47, P15b / P20 consensus), or (ii) X at position 7 is optional and can be any amino acid; X at position 10 is M, E, γ-glutamate, G, A, S, T, or K; and X at position 14 is optional and can be Q, E, γ-glutamate, G, A, or S, TIFF2025096309000059.tif4128 (SEQ ID NO:12, P15 / 20min consensus) relates to an isolated peptide having the amino acid sequence of

[0083] In certain embodiments, these peptides according to SEQ ID NO:47 or 12 also have the structural features defining the peptide of SEQ ID NO:93, in which case there are no methionine and cysteine residues. Thus, in these embodiments, X at position 15 of SEQ ID NO:47 is other than M, and X at position 18 of SEQ ID NO:47 is E, γ-glutamate, G, A, S, T, or K. Similarly, X at position 7 of SEQ ID NO:12 is other than M, and X at position 10 of SEQ ID NO:47 is E, γ-glutamate, G, A, S, T, or K.

[0084] The length of the peptide according to this third aspect is preferably less than about 100 amino acids, or alternatively less than 90 amino acids, less than 80 amino acids, less than 70 amino acids, less than 60 amino acids, or less than about 50 amino acids. In certain embodiments, the peptide is 20 - 44 amino acids in length.

[0085] In certain embodiments, a peptide sharing the structure of SEQ ID NO:47 has at least one of the residues at positions 6 and 22 of SEQ ID NO:47 other than Gln (Q), i.e., E, γ-glutamate, G, A, or S. In certain embodiments, both the residues at positions 6 and 22 of SEQ ID NO:47 are other than Gln (Q), or the residue at position 6 is other than Gln (Q) and the residue at position 22 does not exist.

[0086] Exemplary peptides that share a consensus structure with SEQ ID NO:47 or 12, or are derived from one of SEQ ID NO:47 and 12 and have the consensus structure of SEQ ID NO:93, are identified in Table 3 below.

[0087] (Table 3) Peptide variants of peptide P15 / P20 consensus (SEQ ID NO:47 or 12) The selected peptides in Table 3 of TIFF2025096309000060.tif137169 include solubility tags shown in italics that contain SEEEE. Peptides that include the sequences shown in Table 3 but do not contain this particular solubility tag (or have a different solubility tag) are also contemplated herein.

[0088] In this embodiment, the corresponding wild-type amino acid sequence corresponds to amino acids 52 - 96 of the Pseudomonas syringae HrpW sequence identified in PCT application WO01 / 98501 to Fan et al., which is hereby incorporated by reference in its entirety. For the purpose of comparing the properties of the peptides of the present invention, a polypeptide comprising the amino acid sequence of SEQ ID NO:48, or a peptide consisting of the amino acid sequence of SEQ ID NO:48, is intended to be used as a reference.

[0089] In certain embodiments, the peptide comprises one or more mutations as compared to the corresponding wild-type amino acid sequence of SEQ ID NO:48. These one or more mutations include deletions or substitutions as compared to SEQ ID NO:48. In certain embodiments, the one or more mutations improve the solubility, stability, and / or resistance to chemical degradation of the isolated peptide in an aqueous solution as compared to a polypeptide comprising or consisting of the corresponding wild-type amino acid sequence of SEQ ID NO:48. In this embodiment, the isolated peptide is stable when dissolved in water; as described above, it is resistant to chemical degradation under aqueous conditions in the presence of a pH buffer and a biocide; and / or has a solubility in an aqueous solution of at least about 1.0 mg / ml.

[0090] In certain embodiments, the isolated peptide comprising the amino acid sequence of SEQ ID NO: 47 is a peptide having an overall length of 20 to 36 amino acids, consisting essentially of SEQ ID NO: 49, SEQ ID NO: 63, or SEQ ID NO: 64, and in another embodiment, the isolated peptide consists of SEQ ID NO: 49, SEQ ID NO: 63, or SEQ ID NO: 64.

[0091] In certain embodiments, the isolated peptide comprising the amino acid sequence of SEQ ID NO: 47 is a peptide having an overall length of less than 100 amino acids, and its amino acid sequence comprises SEQ ID NO: 65. In certain embodiments, the amino acid sequence of the peptide consists essentially of SEQ ID NO: 65, and in another embodiment, the isolated peptide consists of SEQ ID NO: 65.

[0092] A further aspect of the present invention is (i) X at position 4 is F or Y; X at position 6 is Q, E, γ-glutamate, G, A, or S; X at position 7 is optional and, according to one embodiment, can be M, E, γ-glutamate, G, A, S, T, or K, or according to another embodiment, can be L; X at position 9 is M, E, γ-glutamate, G, A, S, T, or K; X at position 10 is H or N; X at position 14 is E, γ-glutamate, D, or isoD; X at position 17 is Q, E, γ-glutamate, G, A, or S; and X at position 19 is Q, E, γ-glutamate, G, A, or S, TIFF2025096309000061.tif4128 (SEQ ID NO: 66, P6 / 6a consensus), or (ii) X at position 1 is F or Y; X at position 3 is Q, E, γ-glutamate, G, A, or S; The 4-position X is optional and, according to one embodiment, can be M, E, γ-glutamate, G, A, S, T, or K, or according to another embodiment, can be L; The 6-position X is M, E, γ-glutamate, G, A, S, T, or K; The 7-position X is H or N; and The 11-position X is E, γ-glutamate, D, or iso-D, TIFF2025096309000062.tif4128 (SEQ ID NO:135, P6 / 6a min consensus) relates to an isolated peptide having the amino acid sequence of and containing one or more mutations as compared to the corresponding wild-type amino acid sequence. In certain embodiments, the one or more mutations improve the water solubility, stability, or resistance to chemical degradation of the isolated peptide as compared to a polypeptide containing the corresponding wild-type amino acid sequence.

[0093] The comparable wild-type sequence corresponds to amino acids 85-105 of the full-length harpin of Xanthomonas oryzae pv. oryzae (Ji et al., “Two Coiled-Coil Regions of Xanthomonas oryzae pv. Oryzae Harpin Differ in Oligomerization and Hypersensitive Response Induction,” Amino Acids 40:381-392 (2011), which is hereby incorporated by reference in its entirety). This comparable wild-type sequence is a peptide consisting of the amino acid sequence TIFF2025096309000063.tif5128.

[0094] In certain embodiments, the peptide according to this aspect corresponds to amino acids 85 - 104 of the full-length harpin of Xanthomonas oryzae pv. oryzae (Ji et al., “Two Coiled-Coil Regions of Xanthomonas oryzae pv. Oryzae Harpin Differ in Oligomerization and Hypersensitive Response Induction,” Amino Acids 40:381-392 (2011), which is incorporated herein by reference in its entirety). It does not consist of the amino acid sequence of TIFF2025096309000064.tif5128.

[0095] In certain embodiments, the peptide of this aspect does not include the peptide sequence of motif 2 as described in U.S. Patent No. 8,440,881, as defined as TIFF2025096309000065.tif5141. By way of example, the peptide according to SEQ ID NO:66 does not include a peptide having M / A / T at position 7 when all other alignment residues match the sequence of motif 2; or the peptide according to SEQ ID NO:66 does not include a peptide having H / N at position 10 when all other alignment residues match the sequence of motif 2; or the peptide according to SEQ ID NO:66 does not include a peptide having E / D at position 14 when all other alignment residues match the sequence of motif 2. Similarly, the peptide according to SEQ ID NO:135 does not include a peptide having M / A / T at position 4 when all other alignment residues match the sequence of motif 2; or the peptide according to SEQ ID NO:135 does not include a peptide having H / N at position 7 when all other alignment residues match the sequence of motif 2; or the peptide according to SEQ ID NO:135 does not include a peptide having E / D at position 11 when all other alignment residues match the sequence of motif 2.

[0096] In certain embodiments, the peptide comprises one or more mutations compared to the corresponding wild-type amino acid sequence of SEQ ID NO: 153. These one or more mutations include deletions or substitutions compared to SEQ ID NO: 153. In certain embodiments, the one or more mutations improve the solubility, stability, or resistance to chemical degradation of the isolated peptide in an aqueous solution compared to a polypeptide comprising or consisting of the corresponding wild-type amino acid sequence of SEQ ID NO: 153.

[0097] The length of the peptide according to this aspect is preferably less than about 100 amino acids, or alternatively less than 90 amino acids, less than 80 amino acids, less than 70 amino acids, less than 60 amino acids, or less than about 50 amino acids. In certain embodiments, the peptide is 19 - 50 amino acids in length.

[0098] In certain embodiments, the peptides according to SEQ ID NO: 66 and 135 also have the structural features that define the peptide of SEQ ID NO: 93, in which case there are no methionine and cysteine residues. For example, in the peptide according to SEQ ID NO: 66 that does not contain a methionine amino acid residue, X at position 7 is, if present, E, γ-glutamate, G, A, S, T, K, or L; and X at position 9 is E, γ-glutamate, G, A, S, T, or K. Similarly, in the peptide according to SEQ ID NO: 135 that does not contain a methionine amino acid residue, X at position 4 is, if present, E, γ-glutamate, G, A, S, T, K, or L; and X at position 6 is E, γ-glutamate, G, A, S, T, or K.

[0099] In certain embodiments, a peptide sharing the structure of SEQ ID NO:66 has at least one of the residues at positions 6, 17, and 19 of SEQ ID NO:66 being other than Gln (Q), i.e., E, γ-glutamate, G, A, or S. In certain embodiments, two or three of the residues at positions 6, 17, and 19 of SEQ ID NO:66 are other than Gln (Q). Similarly, in a peptide sharing the structure of SEQ ID NO:135 according to one embodiment, these peptides have the residue at position 6 being other than Gln (Q), i.e., E, γ-glutamate, G, A, or S.

[0100] Exemplary peptides that share a consensus structure with SEQ ID NO:66 or 135, or are derived from one of SEQ ID NO:66 and 135 and have the consensus structure of SEQ ID NO:93, are identified in Table 4 below.

[0101] (Table 4) Peptide variants of the peptide P6 / P6b consensus (SEQ ID NO:66 or 135) The selected peptides in Table 4, TIFF2025096309000066.tif125156, include solubility tags indicated in italics, including SEE, SEEE, and SEEEE. Peptides that include the sequences shown in Table 4 but do not include these specific solubility tags (or have different solubility tags) are also contemplated herein.

[0102] Another aspect of the present invention is (i) X at position 1 can be any amino acid, but preferably Q, S, E, γ-glutamate, A, T, G, D, isoD, N, K, or R; X at position 2 can be any amino acid, but preferably Q, S, E, γ-glutamate, A, T, G, D, isoD, N, K, or R; X at position 3 can be any amino acid, but preferably P, Q, S, E, γ-glutamate, A, T, G, D, isoD, N, K, or R; The X at position 4 can be any amino acid, preferably I, Q, S, E, γ-glutamate, A, T, G, D, N, isoD, K, or R; The X at position 5 can be any amino acid, preferably D, isoD, S, E, γ-glutamate, A, T, G, N, Q, K, or R; The X at position 6 can be any amino acid, preferably R, Q, S, E, γ-glutamate, A, T, G, D, isoD, N, or K; The X at position 7 can be any amino acid, preferably Q, S, E, γ-glutamate, A, T, G, D, isoD, N, K, or R; The X at position 8 can be any amino acid, preferably T, Q, S, E, γ-glutamate, A, G, D, isoD, N, K, or R; The X at position 9 can be any amino acid, preferably I, Q, S, E, γ-glutamate, A, T, G, D, isoD, N, K, or R; The X at position 10 can be any amino acid, preferably E, γ-glutamate, Q, S, A, T, G, D, isoD, N, K, or R; The X at position 11 can be any amino acid, preferably Q, S, E, γ-glutamate, A, T, G, D, isoD, N, K, or R; The X at position 13 can be any amino acid, preferably A, S, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; The X at position 14 can be any amino acid, preferably Q, A, S, T, G, D, isoD, E, γ-glutamate, N, K, or R; The X at position 17 can be any amino acid, preferably A, S, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; The X at position 18 can be any amino acid, preferably Q, A, S, T, G, D, isoD, E, γ-glutamate, N, K, or R; The X at position 21 can be any amino acid, preferably K, A, S, T, G, D, isoD, E, γ-glutamate, Q, N, or R; The 22nd X can be any amino acid, but preferably S, A, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; The 25th X can be any amino acid, but preferably S, A, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; The 26th X can be any amino acid, but preferably P, S, A, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; and The 27th X can be any amino acid, but preferably Q, S, A, T, G, D, isoD, E, γ-glutamate, N, K, or R, TIFF2025096309000067.tif5134 (SEQ ID NO:18, P17 / 18), or (ii) The 2nd X can be any amino acid, but preferably A, S, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; The 3rd X can be any amino acid, but preferably Q, A, S, T, G, D, isoD, E, γ-glutamate, N, K, or R; The 6th X can be any amino acid, but preferably A, S, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; The 7th X can be any amino acid, but preferably Q, A, S, T, G, D, isoD, E, γ-glutamate, N, K, or R; The 10th X can be any amino acid, but preferably K, A, S, T, G, D, isoD, E, γ-glutamate, Q, N, or R; and The 11th X can be any amino acid, preferably S, A, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R, TIFF2025096309000068.tif5128 (SEQ ID NO:25, P17 / 18min consensus) relates to a peptide having the amino acid sequence of.

[0103] In certain embodiments, the peptide comprises one or more mutations as compared to the corresponding wild-type amino acid sequence of Erwinia amylovora HrpW. These one or more mutations include deletions or substitutions as compared to the wild-type HrpW sequence. In certain embodiments, the one or more mutations improve the solubility, stability, and / or resistance to chemical degradation of the isolated peptide in an aqueous solution as compared to a polypeptide comprising or consisting of the corresponding wild-type amino acid sequence of Erwinia amylovora HrpW.

[0104] The PCT application WO01 / 98501 to Fan et al., which is hereby incorporated by reference in its entirety, identifies two hypersensitive response-inducing domains of HrpW Ea The first domain extends from amino acid 5 to amino acid 64 of HrpW, specifically from amino acid 31 to amino acid 57. The second domain extends from amino acid 103 to amino acid 146 of HrpW, specifically from amino acid 116 to amino acid 140. Despite this description in Fan et al., that reference only determines a single peptide fragment of HrpW Ea which is the peptide consisting of amino acids 10 to 59. Ea The comparable wild-type sequence corresponds to amino acids 10 to 59 of the full-length Erwinia amylovora HrpW sequence identified in the PCT application WO01 / 98501 to Fan et al., which is hereby incorporated by reference in its entirety. For the purpose of comparing the properties of the peptides of the present invention, the peptide consisting of amino acids 10 to 59 of Erwinia amylovora HrpW is intended to be used as a reference. Ea

[0105]

[0106] In certain embodiments, the peptide of this aspect has an amino acid sequence corresponding to amino acids 10 to 59 of the full-length Erwinia amylovora HrpW (the PCT application WO01 / 98501 to Fan et al., which is hereby incorporated by reference in its entirety) It does not consist of TIFF2025096309000069.tif5170.

[0107] In certain embodiments, the peptide contains one or more mutations compared to the corresponding wild-type amino acid sequence of SEQ ID NO: 162. These one or more mutations include deletions or substitutions compared to SEQ ID NO: 162. In certain embodiments, the one or more mutations improve the solubility, stability, and / or resistance to chemical degradation of the isolated peptide in an aqueous solution compared to a polypeptide comprising or consisting of the corresponding wild-type amino acid sequence of SEQ ID NO: 162.

[0108] The length of the peptide according to this fourth aspect is preferably less than about 100 amino acids, or alternatively less than 90 amino acids, less than 80 amino acids, less than 70 amino acids, less than 60 amino acids, or less than about 50 amino acids. In certain embodiments, the peptide is 13 - 50 amino acids in length, or still 13 - 40 amino acids in length.

[0109] In certain embodiments, the peptides according to SEQ ID NO: 18 and 25 also have the structural features defining the peptide of SEQ ID NO: 93, in which case there are no methionine and cysteine residues. For example, if the peptide containing SEQ ID NO: 18 does not contain a methionine amino acid residue, the amino acid at position 12 is L. Similarly, if the peptide containing SEQ ID NO: 25 does not contain a methionine amino acid residue, the amino acid at position 1 is L.

[0110] In certain other embodiments, one or more of amino acids 1 to 11 and / or 25 to 27 are not present in the isolated peptide of SEQ ID NO: 18. For example, a peptide that does not contain amino acids 25 to 27 shows an improvement in stability compared to the wild-type sequence.

[0111] Exemplary peptides that share a consensus structure with SEQ ID NO:18 or 25, or are derived from either SEQ ID NO:18 or 25, or have the consensus structure of SEQ ID NO:93, are identified in Table 5 below.

[0112] (Table 5) Peptide variants of the peptide P17 / P18 consensus (SEQ ID NO:18 or 25) TIFF2025096309000070.tif86153In P17, P17a, and the wild-type sequence TIFF2025096309000071.tif4128The selected peptides in Table 5 contain a solubility tag, indicated in italics, that includes SEEEEE. Peptides that contain the sequences shown in Table 5 but do not contain these specific solubility tags (or have different solubility tags) are also contemplated herein.

[0113] In this embodiment, the wild-type amino acid sequence corresponds to amino acids 10 to 59 of the Erwinia amylovora HrpW sequence identified in PCT application WO01 / 98501 to Fan et al., which is hereby incorporated by reference in its entirety. For the purpose of comparing the properties of the peptides of the present invention, a peptide consisting of amino acids 10 to 59 of Erwinia amylovora HrpW is intended to be used as a reference.

[0114] A further aspect of the present invention is X at position 1 is optional and can be L, I, V, F, or M; X at position 3 can be any amino acid, but is preferably K, A, S, T, G, D, isoD, E, γ-glutamate, Q, N, or R; X at position 4 can be any amino acid, but is preferably A, S, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; X at position 7 can be any amino acid, but is preferably K, A, S, T, G, D, isoD, E, γ-glutamate, Q, N, or R; The 10 X's can be any amino acid, but preferably can be A, S, T, G, D, isoD, E, γ-glutamate, Q, N, K, or R; and the 11 X's can be any amino acid, but preferably can be R, A, S, T, G, D, isoD, E, γ-glutamate, Q, N, or K, relates to a peptide having the amino acid sequence of TIFF2025096309000072.tif5128 (SEQ ID NO:26, P19 consensus).

[0115] As noted above, the PCT application WO01 / 98501 to Fan et al., which is hereby incorporated by reference in its entirety, identified two hypersensitive response-inducing domains of HrpW Ea one of which extends from amino acid 103 to amino acid 146 of HrpW, and in particular from amino acid 116 to amino acid 140. Despite this description in Fan et al., this reference did not identify peptide fragments of HrpW Ea that contain this domain. Ea

[0116] A comparable wild-type sequence corresponds to amino acids 116 to 140 of the full-length Erwinia amylovora HrpW sequence identified in the PCT application WO01 / 98501 to Fan et al., which is hereby incorporated by reference in its entirety. For the purpose of comparing the properties of the peptides of the present invention, a peptide consisting of amino acids 116 to 140 of Erwinia amylovora HrpW is intended to be used as a reference.

[0117] In certain embodiments, the peptide of this aspect does not consist of the amino acid sequence corresponding to amino acids 116 to 140 of full-length Erwinia amylovora HrpW (PCT application WO01 / 98501 to Fan et al., which is hereby incorporated by reference in its entirety) TIFF2025096309000073.tif5128.

[0118] ​In certain embodiments, the peptide comprises one or more mutations as compared to the corresponding wild-type amino acid sequence of SEQ ID NO:89. These one or more mutations include deletions or substitutions as compared to SEQ ID NO:89. In certain embodiments, the one or more mutations improve the solubility, stability, and / or resistance to chemical degradation of the isolated peptide in an aqueous solution as compared to a polypeptide comprising or consisting of the corresponding wild-type amino acid sequence of SEQ ID NO:89.

[0119] The length of the peptide according to this aspect is preferably less than about 100 amino acids, or alternatively less than 90 amino acids, less than 80 amino acids, less than 70 amino acids, less than 60 amino acids, or less than about 50 amino acids. In certain embodiments, the peptide is 18 to 50 amino acids in length.

[0120] Exemplary peptides that share a consensus structure with SEQ ID NO:26 or are derived from SEQ ID NO:26 and have the consensus structure of SEQ ID NO:93 are identified in Table 6 below.

[0121] (Table 6) Peptide Variants of Peptide P19 Consensus (SEQ ID NO:26) The selected peptides in Table 6, TIFF2025096309000074.tif62167, contain solubility tags, indicated in italics, that include SEEEE. Peptides that include the sequences shown in Table 6 but do not include this particular solubility tag (or have a different solubility tag) are also contemplated herein.

[0122] Certain peptides in Table 6 also possess the structural features that define the peptide of SEQ ID NO:93, in which case there are no methionine and cysteine residues. When these peptides also meet the limitations of SEQ ID NO:93, amino acid residue 1 of SEQ ID NO:26, if present, is L, I, V, or F; amino acid 5 of SEQ ID NO:26 is L; and amino acids 12 and 13 of SEQ ID NO:26 are independently L, I, V, or F.

[0123] Yet another aspect of the present invention is (i) X at position 1 can be Q, N, D, E, γ-glutamate, isoD, or S; X at position 2 can be D, E, γ-glutamate, isoD; X at position 3 can be P, D, E, isoD, or γ-glutamate; X at position 4 can be M, A, S, D, E, isoD, or γ-glutamate; X at position 5 can be Q, E, or γ-glutamate; X at position 6 can be A, E, or γ-glutamate; X at position 8 can be M, L, E, Q, D, N, G, A, S, isoD, or γ-glutamate; X at position 9 can be Q, N, E, D, G, A, S, isoD, or γ-glutamate; X at position 12 can be Q, N, E, D, G, A, S, isoD, or γ-glutamate; X at position 13 can be Q, N, E, D, G, A, S, isoD, or γ-glutamate; and X at position 16 can be K, Q, N, E, D, R, G, A, or S, TIFF2025096309000075.tif4128 (SEQ ID NO:13, P14d consensus), or (ii) X at position 2 can be M, L, E, Q, D, N, G, A, S, isoD, or γ-glutamate; The X at position 3 can be Q, N, E, D, G, A, S, isoD, or g-glutamate; The X at position 6 can be Q, N, E, D, G, A, S, isoD, or g-glutamate; The X at position 7 can be Q, N, E, D, G, A, S, isoD, or g-glutamate; and The X at position 10 can be K, Q, N, E, D, R, G, A, or S, TIFF2025096309000076.tif4128 (SEQ ID NO:14, P14d min consensus) relates to a peptide having the amino acid sequence of.

[0124] In certain embodiments, the peptide comprises one or more mutations compared to the corresponding wild-type amino acid sequence of Ralstonia solanacearum (formerly Pseudomonas solanacearum) PopA. These one or more mutations include deletions or substitutions compared to the wild-type PopA sequence. In certain embodiments, the one or more mutations improve the solubility, stability, and / or resistance to chemical degradation of the isolated peptide in an aqueous solution compared to a polypeptide comprising or consisting of the corresponding wild-type amino acid sequence of Ralstonia solanacearum PopA.

[0125] The comparable wild-type sequence corresponds to amino acids 92 to 125 of the Ralstonia solanacearum (formerly Pseudomonas solanacearum) PopA sequence identified in PCT application WO01 / 98501 to Fan et al., which is hereby incorporated by reference in its entirety. For the purpose of comparing the properties of the peptides of the present invention, the wild-type peptide of Fan et al. consisting of amino acids 92 to 125 of Ralstonia solanacearum PopA is intended to be used as a reference.

[0126] In certain embodiments, the peptide of this aspect corresponds to amino acids 92 to 125 of Ralstonia solanacearum PopA (see PCT application WO01 / 98501 to Fan et al., which is incorporated herein by reference in its entirety). It does not consist of the amino acid sequence of TIFF2025096309000077.tif5133.

[0127] The length of the peptide according to this aspect is preferably less than about 100 amino acids, or alternatively less than 90 amino acids, less than 80 amino acids, less than 70 amino acids, less than 60 amino acids, or less than about 50 amino acids. In certain embodiments, the peptide is 12 - 50 amino acids in length.

[0128] Exemplary peptides that share a consensus structure with SEQ ID NO:13 or 14, or are derived from SEQ ID NO:13 and have the consensus structure of SEQ ID NO:93, are identified in Table 7 below.

[0129] (Table 7) Peptide variants of peptide P14d (SEQ ID NO:13) The selected peptides in TIFF2025096309000078.tif38167 Table 7 contain a solubility tag, indicated in italics, that includes SEEEEE. Peptides that contain the sequences shown in Table 7 but do not contain this particular solubility tag (or have a different solubility tag) are also contemplated herein.

[0130] Note that the C-terminal lysine residue is thought to be required for HR induction by the p14d variant. This is a minor deviation from the canonical sequence of SEQ ID NO:93. Without being bound by theory, it is thought that the single hydrophilic amino acid (LVKLL) between the two hydrophobic doublet sequences within the p14d variant may render the C-terminal lysine necessary.

[0131] Certain peptides according to this aspect also have the structural features that define the peptide of SEQ ID NO:93, in which case there are no methionine and cysteine residues. For example, in the peptide containing SEQ ID NO:13, amino acid residue 4 of SEQ ID NO:13 is A, S, D, isoD, E, or γ-glutamate, and amino acid residue 8 of SEQ ID NO:13 is L, E, γ-glutamate, Q, D, isoD, N, G, A, or S. Similarly, in the peptide containing SEQ ID NO:14, the amino acid residue at position 2 is L, E, γ-glutamate, Q, D, isoD, N, G, A, or S.

[0132] Another aspect of the present invention is (i) X at position 2 can be Q, N, E, γ-glutamate, D, isoD, T, S, A, or G; X at position 3 can be K, Q, N, E, γ-glutamate, D, isoD, T, S, A, or G; X at position 6 can be K, Q, N, E, γ-glutamate, D, isoD, T, S, A, or G; X at position 9 can be E, γ-glutamate, D, isoD, Q, N, T, S, A, or G; and X at position 10 can be A, G, S, T, E, γ-glutamate, D, isoD, Q, or N, TIFF2025096309000079.tif5128 (SEQ ID NO:16, P25 consensus), or (ii) X at position 2 can be T, S, A, G, D, isoD, E, γ-glutamate, Q, or N; X at position 3 can be G, T, S, A, D, isoD, E, γ-glutamate, Q, or N; X at position 6 can be Q, N, E, γ-glutamate, D, isoD, T, S, A, or G; X at position 7 can be K, Q, N, E, γ-glutamate, D, isoD, T, S, A, or G; X at position 10 can be K, Q, N, E, γ-glutamate, D, isoD, T, S, A, or G; The 13-position X can be E, g-glutamate, D, isoD, Q, N, T, S, A, or G; The 14-position X can be A, G, S, T, E, g-glutamate, D, isoD, Q, or N; and The 16-position V is optional, TIFF2025096309000080.tif5128 (SEQ ID NO:17, P25 consensus) relates to a peptide having the amino acid sequence of.

[0133] In certain embodiments, the peptide comprises one or more mutations compared to the corresponding wild-type amino acid sequence of Ralstonia solanacearum (formerly Pseudomonas solanacearum) PopA. These one or more mutations include deletions or substitutions compared to the wild-type PopA sequence. In certain embodiments, the one or more mutations improve the solubility, stability, and / or resistance to chemical degradation of the isolated peptide in aqueous solution compared to a polypeptide comprising or consisting of the corresponding wild-type amino acid sequence of Ralstonia solanacearum PopA.

[0134] The comparable wild-type sequence corresponds to amino acids 206 to 260 of the Ralstonia solanacearum (formerly Pseudomonas solanacearum) PopA sequence identified as the hypersensitive response domain in the PCT application WO01 / 98501 to Fan et al., which is hereby incorporated by reference in its entirety. For the purpose of comparing the properties of the peptides of the present invention, the wild-type peptide of Fan et al. consisting of amino acids 206 to 260 of Ralstonia solanacearum PopA is intended to be used as a reference.

[0135] In certain embodiments, the peptide of this aspect corresponds to amino acids 206 to 260 of Ralstonia solanacearum PopA (see PCT application WO01 / 98501 to Fan et al., which is hereby incorporated by reference in its entirety). It does not consist of the amino acid sequence of TIFF2025096309000081.tif12128.

[0136] The length of the peptide according to this embodiment is preferably less than about 100 amino acids, or alternatively less than 90 amino acids, less than 80 amino acids, less than 70 amino acids, less than 60 amino acids, or less than about 50 amino acids. In certain embodiments, the peptide is 12 - 50 amino acids in length.

[0137] Exemplary peptides that share a consensus structure with one of SEQ ID NO:16 or 17, or are derived from one of SEQ ID NO:16 or 17 and have the consensus structure of SEQ ID NO:93, are identified in Table 8 below.

[0138] (Table 8) Peptide variants of peptide P2 (SEQ ID NO:180) and P25 (SEQ ID NO:182) TIFF2025096309000082.tif71167 * =N - terminal sequence The selected peptides in Table 8 contain a solubility tag indicated in italics, including SEEEEE. Peptides that contain the sequences shown in Table 8 but do not contain these specific solubility tags (or have different solubility tags) are also contemplated herein.

[0139] It should be noted that some of these derivative peptides in Table 8 contain a repeating LT sequence that is not observed in the wild - type sequence. However, it should be noted that these sequences require a larger hydrophobic sequence to cause a hypersensitivity reaction when compared to SEQ ID NO:93. Without being bound by judgment, this is due to the presence of the amino acid valine instead of leucine in the sequence, and furthermore, there is only a single hydrophilic amino acid between hydrophobic doublets It is considered that TIFF2025096309000084.tif5128 may be the cause. These changes are harmful to HR, but their effects can be reversed by the addition of additional hydrophobic residues at the C-terminus of the peptide (...KIL to...KILEALV or...KILNALV).

[0140] Certain peptides according to this aspect also have the structural features that define the peptide of SEQ ID NO:93, and in this case, there are no methionine and cysteine residues. For example, in the peptide containing SEQ ID NO:16, amino acid residue 5 is L; in the peptide containing SEQ ID NO:17, amino acid residue 9 is L.

[0141] Another aspect of the present invention is (i) X at position 2 can be Q, N, E, γ-glutamate, D, isoD, T, S, A, or G; X at position 3 can be Q, N, E, γ-glutamate, D, isoD, T, S, A, or G; X at position 6 can be K, Q, N, E, γ-glutamate, D, isoD, T, S, A, or G; X at position 9 can be E, γ-glutamate, D, isoD, Q, N, T, S, A, or G; X at position 10 can be A, G, S, T, E, γ-glutamate, D, isoD, Q, or N; X at position 13 can be Q, N, E, γ-glutamate, D, isoD, T, S, A, or G; and X at position 14 can be Q, N, E, γ-glutamate, D, isoD, T, S, A, or G, relates to a peptide having the amino acid sequence of TIFF2025096309000085.tif5128 (SEQ ID NO:15, P3min consensus).

[0142] In certain embodiments, the peptide comprises one or more mutations compared to the corresponding wild-type amino acid sequence of Erwinia amylovora HrpN. These one or more mutations include deletions or substitutions compared to the wild-type HrpN sequence. In certain embodiments, the one or more mutations improve the solubility, stability, and / or resistance to chemical degradation of the isolated peptide in an aqueous solution compared to a polypeptide comprising or consisting of the corresponding wild-type amino acid sequence of Erwinia amylovora HrpN.

[0143] The comparable wild-type sequence corresponds to amino acids 137 to 180 or 150 to 180 of the Erwinia amylovora HrpN sequence identified in U.S. Patent No. 7,132,525 to Wei et al., which is hereby incorporated by reference in its entirety. The HrpN peptide comprising aa137 to 180 was identified as a hypersensitive response-inducing fragment, while the HrpN peptide comprising aa150 to 180 could not be expressed and tested. For the purpose of comparing the properties of the peptides of the present invention, the wild-type peptide of Wei et al. consisting of either amino acids 137 to 180 or 150 to 180 of Erwinia amylovora HrpN is intended to be used as a reference.

[0144] In certain embodiments, the peptide of this aspect corresponds to amino acids 137 to 180 of Erwinia amylovora HrpN (see U.S. Patent No. 7,132,525 to Wei et al., which is hereby incorporated by reference in its entirety). It does not consist of the amino acid sequence of TIFF2025096309000086.tif5168, or the 31-amino acid peptide corresponding to its aa150 to 180.

[0145] The length of the peptide according to this aspect is preferably less than about 100 amino acids, or alternatively less than 90 amino acids, less than 80 amino acids, less than 70 amino acids, less than 60 amino acids, less than about 50 amino acids, less than about 40 amino acids, or less than 30 amino acids. In certain embodiments, the peptide is 12 to 30 amino acids in length.

[0146] Exemplary peptides that share a consensus structure with SEQ ID NO:15 or are derived from SEQ ID NO:15 and have the consensus structure of SEQ ID NO:93 are identified in Table 9 below.

[0147] (Table 9) Peptide variants of peptide P3 consensus (SEQ ID NO:15) The selected peptides in Table 9, TIFF2025096309000087.tif43170, include solubility tags indicated in italics, including SEE, SEEEE, and EEEE. Peptides that include the sequences shown in Table 9 but do not include these specific solubility tags (or have different solubility tags) are also contemplated herein.

[0148] Note that the minimum P3 sequence requires a sequence longer than the minimum HR box sequence of SEQ ID NO:93. Without being bound by theory, this may be due to the presence of two phenylalanine residues within the hydrophobic sequence.

[0149] Certain peptides according to this aspect also have the structural features that define the peptide of SEQ ID NO:93, in which case there are no methionine and cysteine residues. For example, in the peptide containing SEQ ID NO:15, amino acid residues 1, 7, and 12 are L.

[0150] Based on the disclosed consensus sequence (SEQ ID NO:93), it is possible to generate novel peptide sequences with predicted HR activity that are significantly distant from bacterial protein sequences. These peptides may contain hydrophilic residues optimized for maximum solubility and chemical stability. In a preferred embodiment, these hydrophilic residues are glutamate. Lysine and arginine are also possible options, however, a large number of these residues cause toxic reactions in plants.

[0151] In addition to the above-described peptides modeled (and modified) based on sequences naturally present within larger HR-inducing proteins, the present invention also contemplates fully synthetic peptides that incorporate the consensus of SEQ ID NO:93. Ideally, these synthetic peptides contain several strongly hydrophilic amino acids that bridge between the hydrophobic residues specified by SEQ ID NO:93. Exemplary synthetic peptides are listed in Table 10 below. These peptides contain the necessary hydrophobic peptides associated with HR induction. The intervening hydrophilic residues are preferably selected from charged amino acids for maximum solubility. Uncharged amino acids can be used, however, a relatively high proportion of uncharged amino acids can cause the resulting peptides to aggregate in solution and form precipitates or gels. Glutamate is preferred over aspartate for chemical stability. Lysine and arginine still have excellent solubility characteristics, however, polycations have resulted in toxic reactions in test plants. As a result, arginine-rich sequences such as P30-1 (SEQ ID NO:211) should be avoided.

[0152] (Table 10) Other HR box peptides The selected peptides in Table 10 of TIFF2025096309000088.tif97143 contain solubility tags indicated in italics, including SEE, EE, DD, or EEE. Peptides that contain the sequences shown in Table 10 but do not contain these specific solubility tags (or have different solubility tags) are also contemplated herein.

[0153] The isolated peptides of the present invention can also be provided in the form of fusion peptides additionally comprising a second amino acid sequence coupled to the peptides of the present invention via a peptide bond. The second amino acid sequence can be a purification tag that aids in purification but can be removed later, i.e., cleaved from the peptide after recovery, such as poly-histidine (His6-), glutathione-S-transferase (GST-), or maltose binding protein (MBP-). A protease-specific cleavage site or a chemical-specific cleavage site (i.e., in a cleavable linker sequence) can be introduced between the purification tag and the desired peptide. Protease-specific cleavage sites are well-known in the literature and include, but are not limited to, the enterokinase-specific cleavage site (Asp)4-Lys that is cleaved after lysine; the factor Xa-specific cleavage site Ile-(Glu or Asp)-Gly-Arg that is cleaved after arginine; the trypsin-specific cleavage site that is cleaved after Lys and Arg; and the Genenase™ I-specific cleavage site Pro-Gly-Ala-Ala-His-Tyr. Chemicals and their specific cleavage sites include, but are not limited to, cyanogen bromide (CNBr) that cleaves at a methionine (Met) residue; BNPS-skatole that cleaves at a tryptophan (Trp) residue; formic acid that cleaves at an aspartic acid-proline (Asp-Pro) peptide bond; hydroxylamine that cleaves at an asparagine-glycine (Asn-Gly) peptide bond; and 2-nitro-5-thiocyanobenzoic acid (NTCB) that cleaves at a cysteine (Cys) residue (see Crimmins et al., “Chemical Cleavage of Proteins in Solution,” Curr. Protocol. Protein Sci., Chapter 11:Unit 11.4 (2005), which is incorporated herein by reference in its entirety). To use one of these cleavage methods, it may be necessary to remove unwanted cleavage sites from within the desired peptide sequence by mutation.For example, p4-7E-cR (SEQ ID NO:40) has been mutated for compatibility with trypsin: the lysine residue at position 7 has been mutated to glutamate, and a C-terminal arginine has been added to represent the product of theoretical trypsin cleavage. Similarly, p19-5 (SEQ ID NO:168) contains the sequence "NP", which can be cleaved under acidic conditions. The mutation of these residues to "DE" in p19-5a (SEQ ID NO:169) blocks this specific cleavage mechanism. The desired peptide product can be further purified to remove the cleaved purification tag.

[0154] The isolated peptides of the present invention can also be provided in the form of a fusion peptide comprising a number of peptide sequences of the present invention linked together by a linker sequence, which may or may not take the form of a cleavable amino acid sequence of the above kind. Such multimeric fusion proteins may or may not contain a purification tag. In one embodiment, each monomeric sequence can contain a purification tag linked to the peptide of the present invention by a first cleavable peptide sequence; a plurality of monomeric sequences can be linked to adjacent monomeric sequences by a second cleavable peptide sequence. Thus, in the expression of the multimeric fusion protein, i.e., in the expression in a host cell, the recovered fusion protein can be treated with a protease or chemical effective to cleave the second cleavable peptide sequence, thereby releasing the individual monomeric peptide sequences containing the purification tag. In affinity purification, the recovered monomeric peptide sequences can be treated with a protease or chemical effective to cleave the first cleavable peptide sequence, thereby releasing the purification tag from the peptide in question. The latter can be further purified using gel filtration and / or HPLC as follows.

[0155] According to one approach, the peptides of the present invention can be synthesized by standard peptide synthesis operations. These can be carried out, without limitation, on automated solid-phase peptide synthesizers including, but not limited to, Applied Biosystems 431A, 433A synthesizers, and Peptide Technologies Symphony or large-scale Sonata or CEM Liberty automated solid-phase peptide synthesizers, and include both FMOC (9-fluorenylmethyloxy-carbonyl) and tBoc (tert-butyloxy-carbonyl) synthesis protocols. The use of alternative peptide synthesis equipment is also contemplated. Peptides prepared using solid-phase synthesis are recovered in substantially pure form.

[0156] The peptides of the present invention may also be prepared by the use of recombinant expression systems, followed by separation and purification of the recombinantly prepared peptides. Generally, this involves inserting the coding nucleic acid molecule into an expression system in which the molecule is heterologous (i.e., not normally present). One or more desired nucleic acid molecules encoding the peptides of the present invention may be inserted into a vector. The heterologous nucleic acid molecule is inserted into the expression system or vector in the proper sense (5'-3') orientation and in the correct reading frame with respect to the promoter and any other 5' and 3' regulatory molecules.

[0157] Representative nucleotide sequences for expression in bacterial and plant hosts are included in Table 11 below.

[0158] (Table 11) TIFF2025096309000089.tif52170Additional codon-optimized DNA and RNA sequences can be generated with routine skill based on the amino acid sequences encoded and the knowledge of the desired transformed organism recited herein.

[0159] The expression (including transcription and translation) of the peptides or fusion polypeptides of the present invention by the DNA construct may be regulated with respect to the expression level, the tissue types in which the expression occurs, and / or the developmental stage of the expression. To control the expression of the DNA construct, several different heterologous control sequences (e.g., promoters and enhancers) are available. These include constitutive, inducible, and regulatable promoters, as well as promoters and enhancers that control expression in a tissue- or transient-specific manner. Exemplary constitutive promoters include the raspberry E4 promoter (U.S. Patent Nos. 5,783,393 and 5,783,394, each incorporated herein by reference in its entirety), the nopaline synthase (NOS) promoter (Ebert et al., Proc. Natl. Acad. Sci. (U.S.A.) 84:5745-5749 (1987), incorporated herein by reference in its entirety), the octopine synthase (OCS) promoter (carried on the tumor-inducing plasmid of Agrobacterium tumefaciens), caulimovirus promoters such as the cauliflower mosaic virus (CaMV) 19S promoter (Lawton et al., Plant Mol. Biol. 9:315-324 (1987), incorporated herein by reference in its entirety), and the CaMV 35S promoter (Odell et al., Nature 313:810-812 (1985), incorporated herein by reference in its entirety), the sesame mosaic virus 35S-promoter (U.S. Patent No. 5,378,619, incorporated herein by reference in its entirety), a light-inducible promoter derived from the small subunit of ribulose-1,5-bis-phosphate carboxylase (ssRUBISCO), the Adh promoter (Walker et al., Proc. Natl. Acad. Sci. (U.S.A.) 84:6624-6628 (1987), incorporated herein by reference in its entirety), the sucrose synthase promoter (Yang et al., Proc. Natl. Acad. Sci. (U.S.A.) 87:4144-4148 (1990)), the R gene complex promoter (Chandler et al., Plant Cell 1:1175-1183 (1989), which is incorporated herein by reference in its entirety), the chlorophyll a / b binding protein gene promoter, the CsVMV promoter (Verdaguer et al., Plant Mol Biol., 37:1055-1067 (1998), which is incorporated herein by reference in its entirety), and the melon actin promoter (PCT Publication No. WO00 / 56863, which is incorporated herein by reference in its entirety). Exemplary tissue-specific promoters include the tomato E4 and E8 promoters (U.S. Patent No. 5,859,330, which is incorporated herein by reference in its entirety) and the tomato 2AII gene promoter (Van Haaren et al., Plant Mol Bio., 21:625-640 (1993), which is incorporated herein by reference in its entirety).

[0160] In a preferred embodiment, the expression of the DNA construct is under the control of a regulatory sequence derived from a gene whose expression is associated with early seed and / or embryo development. Indeed, in a preferred embodiment, the promoter used is a seed-enhanced promoter. Examples of such promoters include the 5' regulatory region derived from genes such as napin (Kridl et al., Seed Sci. Res. 1:209:219 (1991), which is incorporated herein by reference in its entirety), globulin (Belanger and Kriz, Genet. 129: 863-872 (1991), GenBank accession number L22295, each of which is incorporated herein by reference in its entirety), gamma zein Z 27 (Lopes et al., Mol Gen Genet. 247:603-613 (1995), which is incorporated herein by reference in its entirety), the L3 oleosin promoter (U.S. Patent No. 6,433,252, which is incorporated herein by reference in its entirety), phaseolin (Bustos et al., Plant Cell 1(9):839-853 (1989), which is incorporated herein by reference in its entirety), arcelin 5 (U.S. Patent Application Publication No. 2003 / 0046727, which is incorporated herein by reference in its entirety), soybean 7S promoter, 7Sa promoter (U.S. Patent Application Publication No. 2003 / 0093828, which is incorporated herein by reference in its entirety), soybean 7S αβ conglycinin promoter, 7Sα promoter (Beachy et al., EMBO J. 4:3047 (1985); Schuler et al., Nucleic Acid Res. 10(24):8225-8244 (1982), each of which is incorporated herein by reference in its entirety), soybean trypsin inhibitor (Riggs et al., Plant Cell 1(6):609-621 (1989), which is incorporated herein by reference in its entirety), ACP (Baerson et al., Plant Mol. Biol., 22(2):255-267 (1993)), stearoyl-ACP desaturase (Slocombe et al., Plant Physiol. 104(4):167-176 (1994), which is hereby incorporated by reference in its entirety), the soybean a' subunit of β-conglycinin (Chen et al., Proc. Natl. Acad. Sci. 83:8560-8564 (1986), which is hereby incorporated by reference in its entirety), Vicia faba USP (U.S. Patent Application Publication No. 2003 / 229918, which is hereby incorporated by reference in its entirety), and the Zea mays L3 oleosin promoter (Hong et al., Plant Mol. Biol., 34(3):549-555 (1997), which is hereby incorporated by reference in its entirety) are included.

[0161] For example, nucleic acid molecules encoding the peptides of the present invention can be prepared by the phosphoramidite method and solid-phase synthesis using phosphoramidite building blocks derived from protected 2'-deoxynucleosides. To obtain the desired oligonucleotide, the building blocks are sequentially coupled to the growing oligonucleotide chain in the order required for the sequence of the product. Once chain assembly is complete, the product is released from the solid phase into solution, deprotected, collected, and typically purified using HPLC. The limits of solid-phase synthesis are suitable for preparing oligonucleotides up to about 200 nt in length encoding peptides on a scale of about 65 amino acids or less. The ends of the synthetic oligonucleotides can be designed to include specific restriction enzyme cleavage sites to facilitate ligation of the synthetic oligonucleotides into expression vectors.

[0162] For longer peptides, oligonucleotides can be prepared by solid-phase synthesis and then ligated together using various techniques. Recombinant techniques for making fully synthetic genes are outlined, for example, in Hughes et al., “Chapter Twelve - Gene Synthesis: Methods and Applications,” Methods in Enzymology 498:277-309 (2011), which is incorporated herein by reference in its entirety.

[0163] Once an appropriate expression vector has been selected, the desired nucleic acid sequence is cloned into the vector using standard cloning procedures in the art, as described in Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Springs Laboratory, Cold Springs Harbor, N.Y. (1989), which is incorporated herein by reference in its entirety, or U.S. Patent No. 4,237,224 to Cohen and Boyer. The vector is then introduced into an appropriate host.

[0164] To recombinantly express the peptides of the present invention, various host-vector systems may be utilized. Primarily, the vector system needs to be compatible with the host being used. Host-vector systems include, but are not limited to: bacteria transformed with bacteriophage DNA, plasmid DNA, or cosmid DNA; microorganisms such as yeast containing yeast vectors; mammalian cell systems infected with viruses (e.g., vaccinia virus, adenovirus, etc.); insect cell systems infected with viruses (e.g., baculovirus); and plant cells infected with Agrobacterium. The expression elements of these vectors vary in their strength and specificity. Depending on the host-vector system utilized, any one of several appropriate transcriptional and translational elements can be used to practice this and other aspects of the present invention.

[0165] The purified peptide can be obtained by several methods. The peptide is preferably produced in purified form (preferably at least about 80% or 85% pure, more preferably at least about 90% or 95% pure) by conventional techniques. Depending on whether the recombinant host cell has been engineered to secrete the peptide into the growth medium (see U.S. Patent No. 6,596,509 to Bauer et al., which is hereby incorporated by reference in its entirety), the peptide can be isolated and purified by centrifugation (to separate cell components from the supernatant containing the secreted peptide), followed by successive ammonium sulfate precipitation of the supernatant. The fraction containing the peptide is subjected to gel filtration on a suitably sized dextran or polyacrylamide column to separate the peptide from other proteins. If necessary, the peptide fraction may be further purified by HPLC.

[0166] Alternatively, if the peptide in question is not secreted, it can be isolated from recombinant cells using standard isolation and purification schemes. This involves disruption of the cells (e.g., by sonication, freezing, French press, etc.), followed by recovery of the peptide from the cell debris. The above centrifugation, precipitation, and purification procedures can be used to achieve purification. The above purification tags can be used to streamline this process.

[0167] In certain embodiments, purification is not necessary. If purification is not performed, the cell-free lysate can be recovered after centrifugation to remove cell debris. The resulting cell-free lysate can be treated with heat for a sufficient amount of time to inactivate any native proteases in the recovered fraction, for example, at 100 °C for 10 minutes. If desired, one or more biocides, protease inhibitors, and nonionic surfactants can be introduced into such cell-free preparations (see U.S. Patent Application Publication No. 20100043095 to Wei, which is hereby incorporated by reference in its entirety).

[0168] Once the peptides of the invention are recovered, they can be used to prepare a composition comprising a carrier and one or more additives selected from the group consisting of biocides or biocidal agents, protease inhibitors, nonionic surfactants, fertilizers, herbicides, insecticides, fungicides, nematicides, biological inoculants, plant regulators, and mixtures thereof.

[0169] In certain embodiments, the composition comprises greater than about 1 nM of the peptide, greater than about 10 nM of the peptide, greater than about 20 nM of the peptide, greater than about 30 nM of the peptide, greater than about 40 nM of the peptide, greater than about 50 nM of the peptide, greater than about 60 nM of the peptide, greater than about 70 nM of the peptide, greater than about 80 nM of the peptide, greater than about 90 nM of the peptide, greater than about 100 nM of the peptide, greater than about 150 nM of the peptide, greater than about 200 nM of the peptide, or greater than about 250 nM of the peptide. In certain embodiments, the composition comprises less than about 1 nM of the peptide. For example, certain peptides can be present at a concentration of less than about 2 ng / ml, less than about 1.75 ng / ml, less than about 1.5 ng / ml, less than about 1.25 ng / ml, less than about 1.0 ng / ml, less than about 0.75 ng / ml, less than about 0.5 ng / ml, less than about 0.25 ng / ml, or even less than about 0.1 ng / ml.

[0170] Suitable carriers include water, aqueous solutions optionally containing one or more auxiliary solvents, slurries, and solid carrier particles. Exemplary solid carriers include mineral earths such as silicates, silica gel, talc, kaolin, limestone, lime, chalk, oil clay, loess, clay, dolomite, diatomaceous earth, calcium sulfate, magnesium sulfate, magnesium oxide, crushed synthetic materials, and plant-derived products such as cereal grains, bark particles, wood particles, and nut shell particles, cellulose powder, starch and starch derivatives, and further other monosaccharides, disaccharides, and polysaccharides.

[0171] Suitable fertilizers include, but are not limited to, ammonium sulfate, ammonium phosphate, ammonium nitrate, urea, and combinations thereof.

[0172] Suitable insecticides include, but are not limited to, members of the neonicotinoid group such as imidacloprid, clothianidin, and thiamethoxam; members of the organophosphate group such as chlorpyrifos and malathion; members of the pyrethroid group such as permethrin; other natural insecticides such as nicotine, nornicotine, and pyrethrin; members of the carbamate group such as aldicarb, carbofuran, and carbaryl; members of the macrocyclic lactone group such as various abamectin, avermectin, and ivermectin products; members of the diamide group such as chlorantraniliprole, cyantraniliprole, and flubendiamide; chitin synthesis inhibitors, particularly those of the benzoylurea group such as lufenuron and diflubenzuron; and any combinations thereof including two or more, three or more, or four or more insecticides. Additional insecticides are listed in the Compendium of Pesticide Common Names, which is a database run by Alan Wood and is available in electronic form at the alanwood.net internet site.

[0173] Suitable fungicides include, but are not limited to, members of the strobilurin group such as azoxystrobin, pyraclostrobin, trifloxystrobin, picoxystrobin, and fluoxastrobin; members of the triazole group such as ipconazole, metconazole, tebuconazole, triticonazole, tetraconazole, difenoconazole, flutriafol, propiconazole, and prothioconazole; members of the succinate dehydrogenase group such as carboxin, fluxapyroxad, boscalid, and sedaxane; members of the phenylamide group such as metalaxyl, mefenoxam, benalaxyl, and oxadixyl; members of the phenylpyrrole group such as fluazinam; members of the phthalimide group such as captan; members of the dithiocarbamate group such as mancozeb and thiram; members of the benzimidazole group such as thiabendazole; and any combination thereof containing two or more, three or more, or four or more fungicides. Additional fungicides are listed in the Compendium of Pesticide Common Names, a database maintained by Alan Wood and available in electronic form at the alanwood.net Internet site.

[0174] Suitable nematicides include, but are not limited to, chemicals of the carbamate group such as aldicarb, aldoxycarb, oxamyl, carbofuran, and cleothocarb; and chemicals of the organophosphate group such as thionazin, ethoprophos, fenamiphos, fensulfothion, terbufos, isazofos, and ebufos. Additional nematicides are listed in the Compendium of Pesticide Common Names, a database maintained by Alan Wood and available in electronic form at the alanwood.net Internet site.

[0175] Suitable bactericides include, but are not limited to, those based on dichlorophen and benzyl alcohol hemi formal (Proxel® from ICI, Acticide® RS from Thor Chemie, and Kathon® MK from Rohm & Haas), and isothiazolinone derivatives such as alkyl isothiazolinone and benzisothiazolinone (Acticide® MBS from Thor Chemie; Proxel® GXL from ICI). Additional bactericides are listed in the Compendium of Pesticide Common Names, which is a database run by Alan Wood and is available in electronic form at the alanwood.net Internet site.

[0176] Suitable inoculants include, but are not limited to, cultures of the genus Bradyrhizobium, in particular Bradyrhizobium japonicum (products from BASF Vault®), Bacillus subtilis, Bacillus firmus, Bacillus pumilis, Streptomyces lydicus, cultures of the genus Trichoderma, cultures of the genus Pasteuria, other cultures of rhizobia cells (BASF Nodulator® and Rhizo-Flo®), and any combination thereof containing two or more, three or more, or four or more inoculants.

[0177] Plant regulators are natural or synthetic chemical substances that stimulate or inhibit plant biochemical signaling. These are usually, but not exclusively, recognized by receptors on the cell surface and trigger a reaction cascade in the cell. Suitable plant regulators include, but are not limited to, ethephon; ethylene; salicylic acid; acetylsalicylic acid; jasmonic acid; methyl jasmonate; methyl dihydrojasmonate; chitin; chitosan; abscisic acid; and any auxin compound or inhibitor including, but not limited to, (4-chlorophenoxy)acetic acid, (2,4-dichlorophenoxy)acetic acid, and 2,3,5-triiodobenzoic acid; any cytokinin including, but not limited to, kinetin and zeatin; gibberellin; brassinolide; and any combination thereof including combinations of two or more, three or more, or four or more regulators.

[0178] Other suitable additives include buffering agents, wetting agents, coating agents, and abrasive agents. These substances can be used to facilitate the application of the compositions according to the invention. In addition, the compositions can be applied to plant seeds together with other conventional seed formulations and treatment substances including clays and polysaccharides.

[0179] Compositions or systems used for treating plant seeds comprise one or more of the peptides of the present invention, preferably but not exclusively, one of P1, P4-14S, P6a, P14d, P15a, P18, P19, or P25, in combination with one or more insecticides, or one or more nematicides, one or more fungicides, one or more other inoculants, or one or more plant regulators, which are combinations of a plurality of such agents. Suitable insecticides, nematicides, fungicides, inoculants, and plant regulators for these combined treatments include those identified above. These compositions are provided in the form of a single composition at the time of seed treatment. In contrast, systems used for seed treatment may involve multiple treatments, for example, using a composition containing the peptide in one treatment and a composition containing one or more insecticides, nematicides, fungicides, plant regulators, and / or bactericides in another treatment. In the latter embodiment, both of these treatments are carried out substantially simultaneously, i.e., before or approximately at the time of planting.

[0180] One such example comprises one or more peptides of the present invention, including (but not limited to) one of P1, P4-14S, P6a, P14d, P15a, P18, P19, or P25, in combination with Poncho™ (clothianidin) available from Bayer Crop Science, Poncho™ VOTiVO (clothianidin and Bacillus firmus biological nematicide) available from Bayer Crop Science, and Gaucho™ (imidacloprid) available from Bayer Crop Science.

[0181] Another example involves one or more peptides of the invention, including (but not limited to) one of P1, P4-14S, P6a, P14d, P15a, P18, P19, or P25, in combination with Cruiser™ (thiamethoxam) available from Syngenta, CruiserMaxx™ (thiamethoxam, mefenoxam, and fludioxonil) available from Syngenta, Cruiser Extreme™ (thiamethoxam, mefenoxam, fludioxonil, and azoxystrobin) available from Syngenta, Avicta™ (thiamethoxam and abamectin) available from Syngenta, and Avicta™ Complete (thiamethoxam, abamectin, and Clariva Complete™ containing a biological inoculum of Pasteuria nishizawae - Pn1 available from Syngenta), and Avicta™ Complete Corn (thiamethoxam, mefenoxam, fludioxonil, azoxystrobin, thiabendazole, and abamectin) available from Syngenta.

[0182] Another embodiment involves one or more peptides of the invention, including (but not limited to) one of P1, P4-14S, P6a, P14d, P15a, P18, P19, or P25, in combination with Vault Liquid plus Integral (Bradyrhizobium and Bacillus subtilis strain MBI 600 inoculum) available from BASF, Vault NP (Bradyrhizobium japonicum inoculum) available from BASF, and Subtilex NG (Bacillus subtilis biological inoculum) available from BASF.

[0183] The present invention further relates to a method of conferring disease resistance to plants, enhancing plant growth, controlling pests, conferring biotic or abiotic stress tolerance to plants, and / or modulating plant biochemical signal transduction. These methods include applying an effective amount of the isolated peptide of the present invention or the composition of the present invention to a plant or plant seed or to a location where a plant is growing or is predicted to grow. As a result of such application, the peptide contacts the cells of the plant or the plant seed and induces disease resistance, growth enhancement, tolerance to biotic stress, tolerance to abiotic stress, or a change in biochemical signal transduction in the plant or the plant growing from the plant seed. Alternatively, the seeds themselves recovered from such plants can also be applied to the plants with the peptide or composition of the present invention so as to confer disease resistance to the plants, enhance plant growth, affect insect control, confer tolerance to biotic or abiotic stress, and / or modulate biochemical signal transduction and modulate maturation.

[0184] In these embodiments, it is also possible to select the plants or plant seeds or locations to which the isolated peptides or compositions of the present invention are applied. For example, in plots known to contain a high nematode content, by applying the isolated peptides or compositions of the present invention as described herein, plants or plant seeds grown in such plots, or the plots (locations) can be selectively treated; for plants or plant seeds grown in plots with a low nematode content, such treatment may not be necessary. Similarly, in plots with little irrigation, by applying the isolated peptides or compositions of the present invention as described herein, plants or plant seeds grown in such plots, or the plots (locations) can be selectively treated; for plants or plant seeds grown in plots with appropriate irrigation, such treatment may not be necessary. Similarly, in plots prone to flooding, by applying the isolated peptides or compositions of the present invention as described herein, plants or plant seeds grown in such plots, or the plots (locations) can be selectively treated; for plants or plant seeds grown in plots not prone to flooding, such treatment may not be necessary. As another example of such a selection, in plots prone to insect attack during a specific period of the growth period, by applying the isolated peptides or compositions of the present invention as described herein, plants or plant seeds grown in such plots, or the plots (locations) can be selectively treated; the same plots may not need to be treated during an ineffective period of the growth period, or other plots not prone to such attack may remain untreated. Such a selection step can be carried out when implementing each of the methods of use described herein, namely, methods of conferring disease resistance to plants, enhancing plant growth, controlling pests (including insects and nematodes), conferring biotic or abiotic stress tolerance to plants, and / or modulating plant biochemical signal transduction.

[0185] Instead of applying an isolated peptide or a composition containing the same to a plant or a plant seed in order to confer disease resistance to the plant, to effect plant growth, to control insects, to confer stress resistance, and / or to modulate biochemical signal transduction in a plant or a plant grown from the seed, a transgenic plant or a plant seed can be utilized. When utilizing a transgenic plant, this involves providing a transgenic plant transformed with a DNA molecule encoding the peptide of the present invention, and growing the plant under conditions effective for the DNA molecule to be capable of conferring disease resistance to the plant, of enhancing plant growth, of controlling insects, of conferring tolerance to biotic or abiotic stress, and / or of modulating biochemical signal transduction. Alternatively, a transgenic plant seed transformed with a DNA molecule encoding the peptide of the present invention can be provided and planted in soil. The DNA molecule then expresses the peptide, whereby a plant is propagated from the planted seed under conditions effective for the peptide to confer disease resistance to the transgenic plant, to enhance plant growth, to control insects, to confer tolerance to biotic or abiotic stress, and / or to modulate biochemical signal transduction.

[0186] The present invention further relates to a method for improving drought resistance in an excised portion taken from an ornamental plant, post-harvest disease resistance or post-harvest drought resistance in a fruit or vegetable harvested from a plant, and / or improving the shelf life of fruit or vegetable ripening for a fruit or vegetable harvested from a plant. These methods include applying an effective amount of the isolated peptide of the present invention or the composition according to the present invention to a plant or to the location where the plant is growing. As a result of such application, the peptide contacts the cells of the plant or the plant seeds, and induces drought resistance in an excised portion taken from an ornamental plant, post-harvest disease resistance or post-harvest drought resistance in a fruit or vegetable harvested from a plant, and / or improves the shelf life of fruit or vegetable ripening for a fruit or vegetable harvested from a plant. Alternatively, an effective amount of the isolated peptide of the present invention or the composition according to the present invention can be applied to the harvested fruit or vegetable. As a result of such application, the peptide contacts the cells of the harvested fruit or vegetable, and induces post-harvest disease resistance or post-harvest drought resistance in the treated fruit or vegetable, and / or improves the shelf life of fruit or vegetable ripening for the treated fruit or vegetable.

[0187] Instead of applying an isolated peptide or a composition containing the same to a plant or a plant seed to induce improvement in drought resistance in an excised portion taken from an ornamental plant, post-harvest disease resistance or post-harvest drought resistance in a fruit or vegetable harvested from a plant, and / or improvement in the shelf life of fruit or vegetable ripening for a fruit or vegetable harvested from a plant, a transformed plant or a plant seed can be utilized. When a transformed plant is utilized, this involves providing a transformed plant transformed with a DNA molecule encoding the peptide of the present invention, and growing the plant under conditions effective to enable the DNA molecule to induce drought resistance in an excised portion taken from an ornamental plant, post-harvest disease resistance or post-harvest drought resistance in a fruit or vegetable harvested from the transformed plant, and / or improvement in the shelf life of fruit or vegetable ripening for a fruit or vegetable harvested from the transformed plant. Alternatively, a transformed plant seed transformed with a DNA molecule encoding the peptide of the present invention can be provided and planted in soil. Then, the DNA molecule expresses the peptide, whereby a plant is propagated from the planted seed under conditions effective to enable the DNA molecule to induce drought resistance in an excised portion taken from an ornamental plant, post-harvest disease resistance or post-harvest drought resistance in a fruit or vegetable harvested from the transformed plant, and / or improvement in the shelf life of fruit or vegetable ripening for a fruit or vegetable harvested from the transformed plant.

[0188] In these embodiments, it is also possible to select a transformed plant or plant seed to practice the present invention. For example, in a plot of land known to have a high nematode content, the transformed plant or plant seed can be selectively grown in such a plot of land; in a plot of land with a low nematode content, a non-transformed plant or plant seed can be grown. Similarly, in a plot of land with little irrigation, the transformed plant or plant seed can be selectively grown in such a plot of land; in a plot of land with appropriate irrigation, a non-transformed plant or plant seed can be grown. Similarly, in a plot of land prone to flooding, the transformed plant or plant seed can be grown in such a plot of land; in a plot of land not prone to flooding, a non-transformed plant or plant seed can be grown. As another example of such a selection, in a plot of land susceptible to insect attack during a specific period of the growth period, the transformed plant or plant seed can be grown in such a plot of land; in a plot of land not susceptible to such insect attack, a non-transformed plant or plant seed can be grown. Such a selection step can be carried out when practicing each of the methods of use described herein, namely, methods of conferring disease resistance to plants, enhancing plant growth, controlling pests (including insects and nematodes), conferring biotic or abiotic stress tolerance to plants, and / or modulating plant biochemical signal transduction.

[0189] The present invention further relates to methods for improving the drought resistance in excised parts taken from ornamental plants, the post-harvest disease resistance or post-harvest drought resistance in fruits or vegetables harvested from plants, and / or the improvement of the shelf life of the ripening of fruits or vegetables harvested from plants. These methods include applying an effective amount of the isolated peptide of the present invention or the composition according to the present invention to the plant or to the place where the plant is growing. As a result of such application, the peptide contacts the cells of the plant or the plant seeds and induces an improvement in the drought resistance in excised parts taken from ornamental plants, the post-harvest disease resistance or post-harvest drought resistance in fruits or vegetables harvested from plants, and / or the improvement of the shelf life of the ripening of fruits or vegetables harvested from plants. Alternatively, an effective amount of the isolated peptide of the present invention or the composition according to the present invention can be applied to the harvested fruits or vegetables. As a result of such application, the peptide contacts the cells of the harvested fruits or vegetables and induces an improvement in the post-harvest disease resistance or post-harvest drought resistance in the treated fruits or vegetables, and / or the improvement of the shelf life of the ripening of the treated fruits or vegetables.

[0190] In these embodiments, it is also possible to select the plant, excised part, fruit, vegetable, or place to which the isolated peptide or composition of the present invention is applied. For example, harvested excised parts or fruits or vegetables that are transported over long distances or stored for long periods can be selectively treated by applying the isolated peptide or composition of the present invention as described herein; harvested excised parts or fruits or vegetables that are transported locally and intended to be consumed without substantial storage time can be excluded from such treatment.

[0191] Instead of applying an isolated peptide or a composition containing the same to a plant or a plant seed in order to induce drought resistance to an excised portion taken from an ornamental plant, post-harvest disease resistance or post-harvest drought resistance to a fruit or vegetable harvested from a plant, and / or improvement of the shelf life of the fruit or vegetable maturation for a fruit or vegetable harvested from a plant, a transformed plant or a plant seed can be utilized. When utilizing a transformed plant, this provides a transformed plant transformed with a DNA molecule encoding the peptide of the present invention, and the DNA molecule is effective under conditions effective to induce drought resistance in an excised portion taken from an ornamental plant, post-harvest disease resistance or post-harvest drought resistance to a fruit or vegetable harvested from the transformed plant, and / or improvement of the shelf life of the fruit or vegetable maturation for a fruit or vegetable harvested from the transformed plant, including growing the plant. Alternatively, a transformed plant seed transformed with a DNA molecule encoding the peptide of the present invention can be provided and planted in soil. Then, the DNA molecule expresses the peptide, whereby a plant is propagated from the planted seed under conditions effective to induce drought resistance in an excised portion taken from an ornamental plant, post-harvest disease resistance or post-harvest drought resistance to a fruit or vegetable harvested from the transformed plant, and / or improvement of the shelf life of the fruit or vegetable maturation for a fruit or vegetable harvested from the transformed plant.

[0192] In these embodiments, it is also possible to select a transformed plant or a plant seed for carrying out the present invention. For example, if it is found that the harvested excised portion or fruit or vegetable is intended to be transported over a long distance or stored for a long time after harvest, a transformed plant or a plant seed can be selected for growth; if it is found that the harvested excised portion or fruit or vegetable is intended to be transported locally and / or consumed without a substantial storage period, a non-transformed plant or a plant seed can be selected for growth.

[0193] Suitable plants include dicotyledonous and monocotyledonous plants, including agricultural plants, forestry plants, ornamental plants, and horticultural plants, whether in their natural form or genetically modified form. Exemplary plants include, but are not limited to, alfalfa, apple, apricot, asparagus, avocado, banana, barley, bean, beech (Fagus species), begonia, birch, blackberry, blueberry, cabbage, camphor, canola, carrot, castor bean, cherry, chinquapin, citrus, cocoa bean, coffee, corn, cotton, cucumber, gourd, eucalyptus, fir, flax, fodder beet, fuchsia, garlic, foxglove, grape, peanut, hemp, hop, Japanese cedar, Brassica juncea, jute, lentil, lettuce, linseed, melon, mustard, nectarine, oak, oat, oil palm, rape, olive, onion, paprika, pea, peach, pear, pelargonium, pepper, petunia, pine (Pinus species), plum, poplar (Populus species), pome fruits, potato, rape, raspberry, rice, rubber tree, rye, sorghum, soybean, spinach, spruce, squash, strawberry, sugar beet, sugar cane, sunflower, tea tree, teak, tobacco, tomato, triticale, turfgrass, watermelon, wheat, and willow (Salix species), Arabidopsis thaliana, Saintpaulia, poinsettia, chrysanthemum, carnation, and zinnia.

[0194] Regarding changes in biochemical signal transduction, this includes both the enhancement of certain plant biochemical pathways and the attenuation of certain other plant biochemical pathways. Biochemical signal transduction pathways that can be altered by the present invention include gene expression and protein production, metabolite production, and the production of signaling molecules / secondary metabolites. Exemplary biochemical signal transduction pathways and their alterations include, but are not limited to, induction of nitric oxide production, peroxide production, and other secondary metabolites; agonists of the ethylene signal transduction pathway and induction of ethylene response gene expression (see Dong et al., Plant Phys. 136:3628 - 3638 (2004); Li et al., Planta 239:831 - 46 (2014); Chang et al., PLoS One 10,e0125498 (2015), each incorporated herein by reference in its entirety); agonists of the salicylic acid signal transduction pathway and induction of salicylic acid response gene expression (see Dong et al., Plant J. 20:207 - 215 (1999), incorporated herein by reference in its entirety); agonists of the abscisic acid pathway and induction of abscisic acid response gene expression (see Dong et al., Planta 221: 313 - 327 (2005), incorporated herein by reference in its entirety); agonists of the gibberellin signal transduction pathway and induction of gibberellin response gene expression (see Li et al., Planta 239:831 - 46 (2014), incorporated herein by reference in its entirety); antagonists of jasmonic acid signal transduction and inhibition of jasmonic acid response gene expression (see Dong et al., Plant Phys. 136:3628 - 3638 (2004), incorporated herein by reference in its entirety); induction of protease inhibitor expression (see Laluk and Mengiste, Plant J. 68:480 - 494 (2011); Xia et al., Chin. Sci.See Bull 56: 2351-2358 (2011); induction of reactive oxygen species production in plant tissues; induction of the production of immune-related peptides and antimicrobial peptides, including but not limited to peroxidase, superoxide dismutase, chitinase, and β-1,3-glucanase (Wang et al., J. Agric. Food Chem. 59:12527-12533 (2011), which is incorporated herein by reference in its entirety); and induction of expansin gene expression and production (see Li et al., Planta 239:831-46 (2014), which is incorporated herein by reference in its entirety).

[0195] Regarding disease resistance, absolute immunity against infection cannot be conferred, but the severity of the disease is reduced and the onset of symptoms is delayed. All of the lesion number, lesion size, and the scale of spore formation of the fungal pathogen are decreased. This method of conferring disease resistance has the potential to treat diseases that could not be treated previously, treat diseases that might have been treated separately due to cost as a whole, and avoid the use of infectious agents or environmentally harmful substances.

[0196] The method of conferring pathogen resistance to plants according to the present invention is useful in conferring resistance against a wide variety of pathogens including viruses, bacteria, and fungi. In particular, by the method of the present invention, resistance against the following viruses can be achieved: Tobacco mosaic virus and Tomato mosaic virus. According to the method of the present invention, in particular, resistance to the following bacteria can also be conferred to plants: pathogenic Pseudomonas, pathogenic Erwinia, pathogenic Xanthomonas, and pathogenic Ralstonia. By using the method of the present invention, plants can be made resistant, in particular, to the following fungi: Fusarium and Phytophthora.

[0197] Regarding the use of the peptide or composition of the present invention for promoting plant growth, various forms of promoting or enhancing plant growth can be achieved. This can occur at the initial stage when plant growth starts from seeds or at the later stage of the plant life cycle. For example, plant growth according to the present invention can include higher yields, increased plant vigor, increased seedling vigor (i.e., after germination), increased plant weight, increased biomass, increased number of flowers per plant, higher grain and / or fruit yields, increased number of seeds produced, increased percentage of germinated seeds, increased germination rate, increased plant size, decreased plant height (in wheat), larger biomass, more and larger fruits, earlier fruit coloring, earlier flower buds, earlier fruit and plant maturity, more tillers or side shoots, larger leaves, delayed leaf senescence, increased shoot growth, increased root growth, changes in root / shoot architecture, increased protein content, increased oil content, increased carbohydrate content, increased pigment content, increased chlorophyll content, increased total photosynthesis, increased photosynthetic efficiency, decreased respiration (less O2 utilization), compensation for yield-reducing treatments, increased stem durability (and resistance to stem lodging), increased root durability (and resistance to root lodging), better plant growth under low light conditions, and combinations thereof. As a result, the present invention brings significant economic benefits to growers. For example, early germination and early maturity enable crops to grow in regions where, without them, their growth would be impossible due to short growing seasons in those regions. An increase in the percentage of seed germination leads to an improved crop stand and more efficient use of seeds. Higher yields, increased size, and increased biomass production enable greater income generation from a given plot of land.

[0198] Regarding the use of the peptides or compositions of the present invention for controlling pests (including, but not limited to, insects and nematodes which are biological stressors), such pest control includes preventing the pests from coming into contact with plants to which the peptides or compositions of the present invention are applied, preventing direct damage to the plants by feeding damage, driving the pests away from such plants, killing the pests near such plants, interfering with the larvae of insects that eat such plants, preventing the pests from colonizing the host plants, preventing the colonized insects from releasing phytotoxins, interfering with the attachment of eggs on the host plants, and the like. The present invention also prevents subsequent disease damage to plants resulting from pest infestation.

[0199] The present invention is effective against a wide variety of insects (biotic stressors). The European corn borer is a major pest of corn (dent corn and sweet corn), but also eats over 200 plant species including green beans, kidney beans, and lima beans as well as edible soybeans, pepper, potato, and tomato, and also many weed species. Additional insect larval feeding pests that damage a wide variety of vegetable crops include: beet armyworm, fall armyworm, American tobacco budworm, cabbage looper, diamondback moth, cabbage fly, onion fly, seedcorn maggot, pickleworm (melonworm), pepper maggot, and tomato pinworm. Overall, this group of insect pests is the most economically important group of pests for vegetable production worldwide. The present invention is also effective against nematodes, another group of economically important biotic stressors. The soybean cyst nematode (Heterodera glycines) is a major pest of soybeans. The reniform nematode (Rotylenchulus reniformis) is a major pest of cotton and can similarly parasitize additional crop species, particularly soybeans and corn. Additional nematode pests include root-knot nematodes of the genus Meloidogyne (particularly in cotton, wheat, and barley), cyst nematodes of the genus Heterodera (particularly in soybeans, wheat, and barley), lesion nematodes of the genus Pratylenchus, seed gall nematodes of the genus Anguina (particularly in wheat, barley, and rye), and stem nematodes of the genus Ditylenchus. Other biotic stressors include spiders, weeds, and combinations thereof.

[0200] Regarding the use of the peptides or compositions of the present invention for conferring abiotic stress resistance to plants, such abiotic stresses include any environmental factor having a harmful effect on plant physiology and development. Examples of such environmental stresses include climate-related stresses (e.g., drought, flooding, frost, low temperature, high temperature, excessive light, and insufficient light), air pollution stresses (e.g., carbon dioxide, carbon monoxide, sulfur dioxide, NO x , hydrocarbons, ozone, ultraviolet irradiation, acid rain), chemicals (e.g., insecticides, fungicides, herbicides, heavy metals), nutrient stresses (e.g., excess or deficiency of fertilizers, micronutrients, macronutrients, particularly potassium, nitrogen derivatives, and phosphorus derivatives), and improvement of the healing response to wounds. The use of the peptides of the present invention confers resistance to plants against such forms of environmental stress.

[0201] A further aspect of the present invention relates to the use of the peptides of the present invention as toxicity mitigators in combination with one or more active agents for controlling aquatic weeds in water bodies, as described in U.S. Patent Application Publication No. 20150218099 to Mann, which is hereby incorporated by reference in its entirety (i.e., in one composition or in separate compositions).

[0202] Another aspect of the present invention relates to the use of the peptides of the present invention as plant strengtheners in compositions for application to plants grown under conditions of reduced irrigation, the compositions also including at least one antioxidant and at least one radiation manager, and optionally at least one plant growth regulator, as described in U.S. Patent Application Publication No. 20130116119 to Rees et al., which is hereby incorporated by reference in its entirety.

[0203] The method of the present invention involving the application of the peptide or composition can be carried out via various procedures when treating the whole or a part of a plant, including leaves, stems, roots, vegetative parts (e.g., cuttings), fruits, etc. This may (but not necessarily) include infiltration of the peptide into the plant. Suitable application methods include high-pressure or low-pressure spraying, injection, and leaf abrasion in the vicinity where the peptide is applied. When treating plant seeds, in the application embodiments of the present invention, the protein or polypeptide that is a hypersensitive response elicitor can be applied by low-pressure or high-pressure spraying, coating, dipping (e.g., soaking), or injection. Those skilled in the art can envision other suitable application procedures, provided that the hypersensitive response elicitor polypeptide or protein can be effectively contacted with the plant cells or plant seeds. After being treated with the peptide or composition of the present invention, the seeds can be planted in natural or artificial soil and cultivated using conventional procedures to produce vegetables. After propagating the plant from the seeds treated according to the present invention, the plant can be treated with one or more applications of the peptide or composition of the present invention to confer disease resistance to the plant, enhance plant growth, control insects on the plant, confer biotic or abiotic stress tolerance, improve the drought resistance of the excised parts taken out, confer post-harvest disease resistance or post-harvest drought resistance to the harvested fruits or vegetables, and / or improve the shelf life of the ripening of the harvested fruits or vegetables.

[0204] The peptide or composition of the present invention can be applied to a plant or plant seeds alone according to the present invention or as a mixture with other substances. Alternatively, the peptide or composition can be applied separately to the plant, and other substances can be applied at different times.

[0205] In an alternative embodiment of the invention that includes the use of the transformed plants and transformed seeds, it is not necessary to locally apply the peptides of the invention to the plants or seeds. Instead, transformed plants transformed with a DNA molecule encoding the peptide of the invention are produced by procedures well known in the art. A vector suitable for expression in plants (i.e., including translation and transcription regulatory sequences operable in plants) can be microinjected directly into plant cells by using a micropipette for mechanically transferring recombinant DNA. Crossway, Mol. Gen. Genetics, 202:179-85 (1985), which is hereby incorporated by reference in its entirety. The genetic material may be transferred into plant cells using polyethylene glycol. Krens, et al., Nature, 296:72-74 (1982), which is hereby incorporated by reference in its entirety.

[0206] Another method for transforming plant cells with the gene encoding the peptide of the invention is particle bombardment of the host cell (also known as biolistic transformation). This can be achieved in one of several ways. The first involves propelling inert or biologically active particles into the cells. This technique is disclosed in U.S. Pat. Nos. 4,945,050, 5,036,006, and 5,100,792 to Sanford et al., which are hereby incorporated by reference in their entirety. Generally, this procedure involves propelling inert or biologically active particles into the cells under conditions effective to penetrate the outer surface of the cells and incorporate them therein. When using inert particles, the vector can be introduced into the cells by coating the particles with a vector containing heterologous DNA. Alternatively, the target cells can be surrounded with the vector such that the vector is carried into the cells by the wake of the particles. Biologically active particles (e.g., dried bacterial cells containing the vector and heterologous DNA) can also be propelled into plant cells.

[0207] Another method of introduction is fusion of the protoplast with any of another entity, a minicell, a cell, a lysosome, or another fusible lipid surface body. Fraley, et al., Proc. Natl. Acad. Sci. USA, 79:1859-63 (1982), which is hereby incorporated by reference in its entirety. The DNA molecule may be introduced into plant cells by electroporation. Fromm et al., Proc. Natl. Acad. Sci. USA, 82:5824 (1985), which is hereby incorporated by reference in its entirety. In this technique, plant protoplasts are electroporated in the presence of a plasmid containing the expression cassette. The conductive impulse of high electric field strength reversibly permeabilizes the biological membrane to allow the introduction of the plasmid. The electroporated plant protoplasts reform, divide, and regenerate cell walls.

[0208] Another method of introducing a DNA molecule into a plant cell is to infect the plant cell with Agrobacterium tumefaciens or A. rhizogenes that has been pre-transformed with the gene. Under appropriate conditions known in the art, the transformed plant cells are grown to form shoots or roots and further grow into plants. Generally, this procedure involves inoculating a suspension of bacteria into plant tissue and incubating the tissue on a regeneration medium at 25-28 °C for 48-72 hours without using antibiotics. Agrobacterium is a representative genus of the gram-negative family Rhizobiaceae. That genus is the cause of crown gall (A. tumefaciens) and hairy root disease (A. rhizogenes). Plant cells in crown gall tumors and hairy root disease are induced to produce amino acid derivatives known as opines, which are degraded only by the bacteria. The bacterial genes responsible for opine expression are a convenient source of control elements for chimeric expression cassettes. In addition, assays for the presence of opines can be used to identify transformed tissue. Heterologous genetic sequences can be introduced into appropriate plant cells using the Ti plasmid of A. tumefaciens or the Ri plasmid of A. rhizogenes. Upon infection with Agrobacterium, the Ti or Ri plasmid is transferred to the plant cell and stably integrated into the plant genome. J. Schell, Science, 237:1176-83 (1987), which is hereby incorporated by reference in its entirety.

[0209] After transformation, it is necessary to regenerate the transformed plant cells. The regeneration of plants from cultured protoplasts is described in Evans et al., Handbook of Plant Cell Cultures, Vol. 1: (MacMillan Publishing Co., New York, 1983); and Nasil I.R. (ed.), Cell Culture and Somatic Cell Genetics of Plants, Acad. Press, Orlando, Vol. 1, 1984, and Vol. Ill (1986), which are hereby incorporated by reference in their entirety.

[0210] It is known that practically all plants can be regenerated from cultured cells or tissues. The means of regeneration vary for each plant species, but generally, a suspension of transformed protoplasts or a Petri dish containing the transformed explant is first obtained. Callus tissue is formed, shoots can be induced from the callus, and then they can root. Alternatively, embryogenesis can be induced in the callus tissue. These embryos germinate as natural embryos to form plants. The culture medium generally contains various amino acids and hormones such as auxins and cytokinins. In particular, for species such as maize and alfalfa, it is also advantageous to add glutamic acid and proline to the medium. Efficient regeneration depends on the medium, genotype, and culture history. By controlling these three variables, regeneration is usually reproducible and repeatable.

[0211] After stably integrating the expression cassette into the transformed plant, it can be transferred to other plants by sexual crossing. Depending on the species to be crossed, any of several standard breeding techniques can be used.

[0212] Once a transgenic plant of this kind is produced, it can be cultivated according to conventional procedures in the presence of a gene encoding a hypersensitive response elicitor that results in disease resistance, enhanced plant growth, control of insects in plants, abiotic or biotic stress tolerance, improved desiccation resistance of excised parts removed, post-harvest disease resistance or post-harvest desiccation resistance in harvested fruits or vegetables, and / or improved fruit or vegetable maturity life for harvested fruits or vegetables.

[0213] Alternatively, transgenic seeds are recovered from the transgenic plant. These seeds can then be planted in soil and cultivated using conventional procedures to produce transgenic plants. The transgenic plants are propagated from the planted transgenic seeds under conditions effective to confer disease resistance to the plants, enhance plant growth, control insects, confer abiotic or biotic stress tolerance, improve desiccation resistance of excised parts removed, confer post-harvest disease resistance or post-harvest desiccation resistance in harvested fruits or vegetables, and / or confer improved fruit or vegetable maturity life for harvested fruits or vegetables.

[0214] When transgenic plants and plant seeds are used according to the present invention, they can additionally be treated with the same substances used to treat plants and seeds to which the peptide or composition of the present invention is applied. These other substances containing the peptide or composition of the present invention can be applied to the transgenic plants and plant seeds by the above-described procedures including high-pressure or low-pressure spraying, injection, coating, and dipping. Similarly, after the plants are propagated from the transgenic plant seeds, the plants can be treated with one or more applications of the peptide or composition of the present invention to confer disease resistance, enhanced growth, control of insects, abiotic or biotic stress tolerance, desiccation resistance of excised parts removed, post-harvest disease resistance or post-harvest desiccation resistance in harvested fruits or vegetables, and / or improved fruit or vegetable maturity life for harvested fruits or vegetables.

[0215] As described above, such transgenic plants may be treated with conventional plant treatment agents such as fungicides or biocides, protease inhibitors, nonionic surfactants, fertilizers, herbicides, insecticides, fungicides, nematicides, biological inoculants, plant regulators, and mixtures thereof.

Example

[0216] The following examples are provided to illustrate embodiments of the present invention and are in no way intended to limit the scope of the present invention.

[0217] Example 1 Development of the "HR box" peptide of SEQ ID NO:93 The HR box was initially developed based on the examination of several hypersensitivity response-inducing sequences (in particular, P1, SEQ ID NO:4; P4, SEQ ID NO:5; and P15, SEQ ID NO:64). Repeated sequences of leucine and isoleucine residues were identified. P4 was selected as a representative sequence as a basis for mutagenesis studies that should clarify the determinants of HR induction. HR in tobacco was tested as described in Wei, Science 257:85-88 (1992), which is hereby incorporated by reference in its entirety. Briefly, the peptide was dissolved in aqueous solution at a concentration of 500 μg / ml. Serial dilutions four times were performed using an equal volume of water to generate peptide samples in 500, 250, 125, 62.5, 31.25 μg / ml peptide solutions. Nicotiana tabacum cultivar xanthi plants at 5-7 weeks of age (before flowering) were used. Leaves were gently pricked with a needle in a panel of middle leaves. Then, the peptide solution was injected into the wound through a needleless syringe to fill the panel. Each peptide sample was injected into the leaves of two different plants. The leaves were observed and scored for wilting and browning, which are typical lesions of programmed cell death, over the subsequent 48 hours. These mutagenesis studies had three main objectives: (1) increasing the solution stability of the peptide; (2) generating disruptive mutations to identify the residues most important for HR induction; and (3) generating conservative mutations to identify the degree of specificity for specific amino acids.

[0218] The peptide was evaluated for one or more of solubility, stability to chemical degradation, the effect of excipients on solution stability, protection against oxidation, and solution stability studies.

[0219] In deionized water, a 0.2% AI (active ingredient) solution of a chemically synthesized pure peptide was prepared, and the solubility was evaluated by observing the solution for 48 hours at room temperature for evidence of precipitation. P1 (SEQ ID NO:4) was mainly insoluble in water. However, variants containing several glutamine residues instead of glutamate residues (P1-2E, 8E, 11E, 15E, 18E, SEQ ID NO:46) were soluble. P4 (SEQ ID NO:5) and P4-14S (SEQ ID NO:6) were also completely soluble.

[0220] To better quantify peptide solubility, subsequent experiments were performed. 20 - 50 mg of pure peptide was mixed with 0.25 ml of water, and incremental amounts of water were added until the peptide dissolved. These experiments estimated the solubility of P1 (SEQ ID NO:4) to be <1 mg / ml, P4 (SEQ ID NO:5) to be 100 mg / ml, and P1-18K (SEQ ID NO:45) to be 20 mg / ml.

[0221] A 0.2% AI solution of a chemically synthesized pure peptide was prepared in deionized water, 0.25% weight / volume Proxel® GXL (insecticide), and eight 50 millimolar (mM) buffers as follows (separately) to evaluate stability against chemical degradation in various pH buffers: MES pH 5.6, MOPS pH 6.5, citrate pH 7.2, EDDS pH 7.3, EDTA pH 8, phosphate pH 8, imidazole pH 8, and TES pH 8. The solutions were observed by HPLC over several weeks at high temperature (50 °C) for evidence of degradation (percent decrease in peptide signal over time compared to the 0-hour sample). P1-2E, -8E, -11E, -15E, -18E (SEQ ID NO: 46) were more stable than P1 (SEQ ID NO: 4) (over 80% for 40 days compared to 20 days), and P4-14S (SEQ ID NO: 6) was significantly more stable than P4 (SEQ ID NO: 5) (over 80% for 35 days compared to 3 days). The best buffers for P1 and P4-14S were TES pH 8 and citrate pH 7.2, respectively. After several days, precipitation of P1 was observed. The other peptides (P1-2E, -8E, -11E, -15E, -18E; P4, and P4-14s) remained in solution.

[0222] In 50 mM TES pH 8.0 solution in water and trehalose, maltrin, sucrose, or talc (separate formulations) as 20% by weight of the filler with respect to the volume of the solution, an AI solution of 0.2% of a chemically synthesized pure peptide was prepared to evaluate the effect of the filler on the chemical decomposition of P1 and P1-2E, -8E, -11E, -15E, -18E. These solutions were observed by HPLC for evidence of decomposition (decrease in peptide signal over time compared to the sample at time zero) at a high temperature (50 °C) over time. In all mixtures, the concentration of P1 decreased to less than 60% of the original peptide concentration after 6 days of incubation. In contrast, in all samples, the concentrations of p1-2E, -8E, -11E, -15E, -18E remained above 80% of the original concentration for at least 14 days. The best filler for P1-2E, -8E, -11E, -15E, -18E is talc powder (over 80% for 44 days).

[0223] A solution stability study was conducted by preparing an AI solution of 0.2% of a chemically synthesized pure peptide in deionized water, 50 mM TES buffer, 0.25% Proxel GXL, and 30% isopropanol. The peptide solution was analyzed by HPLC for the percentage decrease in peptide signal over time compared to the sample at time zero. The maximum lifespan of P1-2E, -8E, -11E, -15E, -18E is over 80% for 45 days. The maximum lifespan of P4-14S is over 80% for 140 days.

[0224] Solution stability variants: By selecting a peptide sequence (P4, SEQ ID NO:5) that does not contain methionine residues, the solution stability was increased. However, this peptide contained cysteine residues that resulted in very poor stability. Mutation of this cysteine to a conservative substitution serine (a change from sulfur to oxygen in the chemical structural formula) resulted in P4-14s (SEQ ID NO:6), which retained its ability to induce HR. It was later shown (as described above) that P4-14s is a highly stable peptide. In additional studies, one or more glutamine residues were replaced with glutamate residues to reduce the possibility of deamidation in solution. In particular, variants of P1 named P1-2E, 8E, 11E, 15E, 18E (SEQ ID NO:46) contained these mutations at positions 2, 8, 11, 15, and 18. This peptide showed improvement in both solubility and stability when compared to P1.

[0225] Based on the stable backbone of P4-14s (SEQ ID NO:6), a single disruptive mutation was incorporated into specific residues within the sequence. In the case of leucine residues, these were mutated to alanine (a smaller and less hydrophobic side chain, a moderate disruptive mutation) and / or aspartic acid (a side chain with a negative charge, a high disruptive mutation). Depending on the identity of the corresponding amino acid, the intervening sequence was mutated to have a negative charge (aspartic acid or glutamic acid), a hydrophobic side chain (valine), a minimal side chain (alanine), or a small polar side chain (serine). These mutant peptides were tested for the induction of hypersensitivity reactions. Based on the initial HR results, additional mutations were selected. In addition, the spacing between leucine / isoleucine residues was evaluated by deleting a single residue between leucine repeats (denoted as "del") or inserting an alanine residue between leucine repeats (denoted as iA).

[0226] For amino acids important for HR induction, more conservative mutations were selected to determine the specificity of the interaction. Leucine residues were mutated to isoleucine, valine, phenylalanine, or tyrosine, with the latter two residues being less conserved. As described above, these mutants were tested for HR induction.

[0227] The results of these mutagenesis studies are summarized in Table 12 below.

[0228] (Table 12) Summary of Mutations and HR Induction Results TIFF2025096309000090.tif87166

[0229] In Table 12, the sequence of P4-14s is shown along with all the mutations tested at each position. Mutations that did not interfere with the hypersensitive response are labeled "HR-positive mutations" and listed. Mutations that resulted in a reduction in the severity of the hypersensitive response are shown in the rows labeled "weak HR mutations". Mutations that eliminated the hypersensitive response are shown in the red rows labeled "HR-negative mutations". The notations dN2 and dN4 indicate deletions of 2 or 4 residues, respectively, from the start of the peptide; dC2 indicates a deletion of 2 residues from the end of the peptide; del indicates a deletion of the residue at that position; and iA represents an insertion of an alanine residue before that position.

[0230] Example 2 Solubility and Stability of P1 and Mutant Peptides As described above, the P1 and the P1-derived sequences mutated at position 18 (where methionine is replaced by alanine, threonine, or lysine) were evaluated for solution stability and chemical compatibility over 14 days. It should be noted that P1 showed solubility problems at relatively low pH (in deionized aqueous solution, in 50 mM citrate pH 5.6, and in 50 mM MES pH 6.0). In these cases, the peptide concentration increased after 24 hours of incubation at 50 °C. It should be noted that the mutant peptides generally did not show this problem. As shown in Figures 1-3, the data were normalized to 100% peptide at the day 1 time point (peptide 1 in the description) * is shown as, and the data for the original peptide 1 are double asterisks **(marked with). In the stability test dissolved in water (Figure 1), Peptide 1 was moderately stable but showed solubility problems. The 18K and 18A mutants showed slightly higher stability (10 - 25% after 14 days). When dissolved in slightly acidic citrate buffer (Figure 2), P1 showed problems with both solubility and stability. It was not detected by HPLC after 14 days in solution. In contrast, the 18T and 18K mutants retained 80% of the original concentration, and the 18A mutant retained approximately 60% of the original concentration. As shown in Figure 3, at 50 mM MES pH 6.0, P1 showed stronger solubility problems with a 50% increase in the dissolved concentration after 24 hours of incubation at 50 °C. However, it showed better stability than the mutants (10 - 30% after 14 days). In citrate pH 7.2 (Figure 6), P1 showed no solubility problems but insufficient stability (20% of the original concentration after 7 days at 50 °C). In contrast, the 18K and 18T mutants showed >60% stability after 14 days. At 50 mM EDDS, pH 7.3 (Figure 7), Peptide 1 showed insufficient stability, and only 10% of the substance remained after 7 days. In comparison, the mutants retained at least 50% of the starting material after 14 days. At 50 mM imidazole, pH 8.0 (Figure 8), Peptide 1 showed particularly insufficient stability and decreased to less than 10% of the original concentration after only 3 days. In comparison, all mutants showed higher stability, and the 18K and 18T mutants retained 60 - 75% of the original substance after 14 days. Peptide 1 showed relatively good stability in a 50 mM EDTA, pH 8.0 solution (Figure 9), consistent with the performance of the 18A mutant. However, the 18K and 18T mutants showed better stability after 14 days of incubation at 50 °C (15 - 20% improvement). When dissolved in phosphate, pH 8.0 (Figure 10), Peptide 1 is thought to perform better than the stability of the mutants, but it shows solubility problems (some turbidity in the solution). In a 50 mM TES, pH 8.0 solution (Figure 11), more than 90% of Peptide 1 decomposed after 1 week of incubation at 80 °C.In comparison, the 18T, 18K, and 18A variants showed better performance (71%, 58%, and 47% remaining after 14 days of incubation at 50°C).

[0231] Overall, Peptide 1 shows either solubility problems or insufficient stability in a wide variety of buffer solutions. This is addressed by mutating the methionine to other residues. Bulky residues (threonine and lysine) are generally considered more favorable for stability than alanine.

[0232] Example 3 Solubility and Stability of P4 and Mutant Peptides As described above, P4 and P4-derived sequences mutated at position 14 (replacing cysteine with alanine / A, aspartic acid / D, lysine / K, glutamine / Q, and serine / S) were evaluated for solution stability and chemical compatibility over 14 days. Overall, Peptide 4 showed very poor stability due to the presence of cysteine (Figures 12 - 21). After less than 1 day, the original P4 HPLC peak was not detected in the sample. In comparison, all variants showed better stability. Many of these retained at least 50% of the original substance over 14 days at 50°C. Overall, the Peptide 4 variants show better stability at higher pH values (>7.0). It should be noted that p4-14s can usually show 90% stability after 14 days, depending on the conditions. All mutant peptides showed a hypersensitive reaction when infiltrated into tobacco leaves (as in Example 1).

[0233] Example 4 Comparison of the Stability of Peptide 1 and Peptide 4 Peptides 1 and 4 showed a high degree of sequence similarity, but the stabilized variants of peptide 4 were more stable than the p1 variants. To confer the same stability as p4-14s, a series of mutations of p1 were generated. These are p1-1S (SEQ ID NO:109, Table 1), p1-14S (SEQ ID NO:110, Table 1), p1-18Q (SEQ ID NO:115, Table 1), p1-23P (SEQ ID NO:118, Table 1). These peptides, together with p1 and p4-14s, were dissolved in 30% isopropanol, 5 mM DTPA, and 50 mM TES pH 8.0 and tested for stability at 50°C. Similar stability was observed for p4-14S and p1-1S, indicating that the N-terminal amino acid has a strong effect on peptide stability.

[0234] Example 5 Solubility of P15b and Variants Initial results suggested that p15b has solubility problems. It has a relatively high hydrophobicity (0.19). At 0.2% w / v, it is partially soluble in water and insoluble in 50 mM citrate pH ~5.2, citrate pH 7.0, phosphate pH 7.0 (checked), TES pH 8.0, EDTA pH 8.0, and EDDS pH 7.0 respectively. It was at least partially soluble in 50 mM MES pH 6.0 and MOPS pH 6.5. However, P15a dissolved more readily in aqueous solution. Its solubility was >10 mg / ml at 50 mM TES pH 8.0. Additional p15 variants containing poly-glutamate solubility tags were synthesized (p15-59G and p15-59, SEQ ID NO:149 and 150 respectively). When p15-59 was dissolved in 50 mM TES, pH 8.0, it showed a solubility >10 mg / ml (1% w / v).

[0235] Example 6 Stability of P17 / P18 and Variants As described above, P18 (SEQ ID NO:83) was tested for stability and chemical compatibility using various pH buffers. P18 shows relatively poor stability in aqueous buffers at 50°C. Many samples decomposed to 20% of their original concentration within 3 days. One exception was the 50 mM EDTA solution, which decomposed to 35% after 7 days (Figure 24). The mutation of methionine at position 12 to leucine (P18-4, in SEQ ID NO:164) results in moderate stabilization: 60% stability after 14 days. Notably, the truncation of the last 3 amino acids from the C-terminus (P18-1, SEQ ID NO:163) also results in a dramatic increase in stability (>90% stability over a 14-day test).

[0236] Example 7 Stability of P19 and Variants Overall, P19 (SEQ ID NO:89) shows relatively high stability, >80% stability, at 50°C over 14 days under various conditions. The exceptions were the peptides dissolved in water alone (52%) or 50 mM TES pH 8.0 (62%). The mutation of one methionine residue at position 12 to leucine (P19-20L, SEQ ID NO:90) results in a moderate increase in stability when dissolved in 50 mM citrate pH 7.2 or 50 mM TES, pH 8.0 (Figures 25 and 26). When using buffers at lower pH (5.5 - 7.0), the performance of P19 and P19-20L was observed to be similar; over 80% of the peptide was retained over 14 days.

[0237] Example 8 Stability of P14d, P14e, P14f The P14d sequence (SEQ ID NO: 175) is derived from the popA sequence of Ralstonia solanacearum. It matches the HR box motif and induces HR in tobacco leaves. Mutations of methionine residues led to the stabilized peptides P14e (SEQ ID NO: 176) and P14f (SEQ ID NO: 177). The mutant peptides show >85% stability at 50 °C for >50 days (in 50 mM TES, pH 8.0 and 30% isopropanol). During the same period, P14d shows approximately 50% chemical stability.

[0238] Example 9 Growth Test For the growth test, corn and soybean seeds were planted in flats in a greenhouse facility at two seeds per cell. The seeds were germinated and the smaller plants were pinched off, leaving one plant per cell. When the first true leaf was fully open and the second leaf began to open, the plants were first measured for height. This was done by stretching the tallest leaf upward and measuring the distance to the soil. Peptides were dissolved in water at the indicated concentrations (below). The plants were then treated by foliar spraying using a widely available spray bottle until the liquid dripped from the leaves. Four flats of 14 plants each were treated per condition (peptide or control). Corn and soybeans were treated as specified in Table 13 and compared to the matched water-treated control plants. The plants were grown for 14 days. The height of the plants was measured again and growth was quantified as compared to the original height. In some cases, the plants were grown without watering for 2 - 4 days until signs of wilt and drought stress began. At this point, the above-ground parts of the plants were harvested and weighed to measure the fresh mass. Finally, the above-ground material was dried at 70 °C for 48 hours and weighed to measure the dry biomass. The results of these growth tests are shown in Table 13. Growth, dry biomass, and fresh mass were calculated as percent increase over the water-treated control.

[0239] (Table 13) Growth Test Results TIFF2025096309000091.tif73165N.D. = Not measured.

[0240] Among several of the tested peptides, growth and / or an increase in biomass are shown in maize and soybean. It should be noted that P15b did not result in an obvious growth or dry biomass phenotype, but this led to an increase in fresh biomass, which suggests an increase in water uptake or retention. This is an indicator of drought tolerance in those treated plants. Another peptide, P30-3, was observed to result in an increase in growth, fresh mass, and dry biomass.

[0241] Example 10 Minimum sequence required for the HR reaction After determining the most important residues for the induction of the hypersensitive reaction, the inventors designed additional mutations to identify the minimal peptide sequence responsible for this behavior. Due to the hydrophobic nature of the nuclear HR sequence (containing 7 leucine or isoleucine residues out of a total of 13 residues), solubility was recognized as an issue for the minimal peptides. As a result, hydrophilic sequences were added to many of the peptides to bring the hydrophobicity on the Kyte-Doolittle scale to approximately -0.2 for the peptides.

[0242] First, P4 (SEQ ID NO: 35, 36, 38, 39) having a poly-lysine or poly-arginine sequence, i.e., an N-linked polyR or polyK sequence, or a C-linked polyR or polyK sequence was used. However, when infiltrated into tobacco leaves, these peptides resulted in necrotic lesions that are not typical of HR. This led to the hypothesis that poly-cationic sequences result in a toxic reaction when infiltrated into tobacco leaves. Similar necrotic lesions were observed when only poly-lysine and poly-arginine were infiltrated into the leaves. As a result, testing of peptides containing cationic solubility-enhancing sequences was discontinued. It should be noted that HR+ peptides can contain at least one or two cationic amino acids, but a larger number of positive charges are thought to be detrimental. As an alternative to cationic peptides, polyanions, specifically poly-glutamate, were considered. Aspartate has a high likelihood of isomerizing to iso-aspartate, so poly-glutamate was selected and serine was added to the N-terminus to remove the formation of pyroglutamic acid at the N-terminus of the peptide. It is also reasonable to add a glutamate residue to the C-terminus of the peptide.

[0243] The hypersensitivity reaction test was conducted as described in Example #1. For P4, the minimal variant peptide that induced HR was P4-polyE-min3 (SEQ ID NO:33). For P1, the minimal variant peptide that induced HR was P1-polyE-min3 (SEQ ID NO:141). For P18, the minimal variant peptide that induced HR was P18-7 (SEQ ID NO:167). For P19, the minimal variant peptide that induced HR was P19-8 (SEQ ID NO:173). For P15, the minimal variant peptide that induced HR was P15-59 (SEQ ID NO:150). For P14d, the minimal variant peptide that induced HR was P14-30 (SEQ ID NO:178). For P25, the minimal variant peptide that induced HR was P25-11 (SEQ ID NO:188). In addition, to increase solubility, leucine repeat sequences and glutamate residues characteristic of the HR box were incorporated at variable positions to generate minimal peptide sequences. These sequences are P30-2 is TIFF2025096309000092.tif12160. These minimal HR box sequences were soluble at >5 mg / ml in 50 mM TES and generated an HR response when infiltrated into tobacco leaves.

[0244] Similarly, based on the hydrophobic backbone sequences of P3, P25, P14, P15, and P19, additional peptides were developed to enhance solubility. These are listed in Table 10 above.

[0245] Based on the previously described behavior of harpins and HR+ peptides, as described in PCT application WO01 / 98501 to Fan et al., which is hereby incorporated by reference in its entirety, these new peptides are predicted to have a wide range of biological activities including induction of resistance to TMV, resistance to nematodes, increased stress and drought resistance, increased growth, and increased yield.

[0246] Example 11 Derivatives of Peptide P1 that Elicit an HR Response in Tobacco To determine the minimal sequence required for HR and identify important residues, the HR assay (described in Example 1) was performed on variants of P1. The following peptides in Table 14 were determined to be positive for HR.

[0247] (Table 14) TIFF2025096309000093.tif74146

[0248] Example 12 Derivatives of Peptide P3 that Elicit an HR Response in Tobacco To determine the minimal sequence required for HR and identify important residues, the HR assay (described in Example 1) was performed on variants of P3. The following peptides in Table 15 were determined to be positive for HR.

[0249] (Table 15) TIFF2025096309000094.tif39169

[0250] Note that for efficient HR induction, P3 is thought to require a sequence longer than the minimal HR box repeat. This may be due to suboptimal phenylalanine residues and the presence of only one K residue to separate hydrophobic residues present in this sequence ( TIFF2025096309000095.tif4128 and its variants). However, considering that P3-6 and P3-7 elicit HR, it is important to note that additional hydrophobic residues are not strictly necessary.

[0251] Example 13 Derivatives of Peptide P25 that Elicit an HR Response in Tobacco To determine the minimal sequence required for HR and identify important residues, the HR assay (described in Example 1) was performed on variants of P25. The following peptides in Table 16 were determined to be positive for HR.

[0252] (Table 16) TIFF2025096309000096.tif57154

[0253] It is important to note that for HR induction, the P25 variant is thought to require more sequence than the minimal HR sequence (SEQ ID NO:93). This may be due to the presence of valine residues in preferred locations for leucine, or the presence of one hydrophilic residue between hydrophobic repeats. TIFF2025096309000097.tif5128 may be the cause. The inventors included P25-15, P25-16, and P25-17 as HR+, but they showed a very weak hypersensitive response that occurred only in some tobacco plants even at the highest application rates. Notably, as suggested by the biological response to P25-15, the additional sequence portion is thought not to require leucine / isoleucine / valine residues.

[0254] Example 14 Derivatives of Peptide P14d that Elicit an HR Response in Tobacco To determine the minimal sequence required for HR and identify important residues, the HR assay (described in Example 1) was performed with variants of P14. The following peptides in Table 17 were determined to be positive for HR.

[0255] (Table 17) TIFF2025096309000098.tif39163

[0256] It is important to note that for HR induction, the P14d variant is thought to require more sequence than the minimal HR sequence (SEQ ID NO:93). In particular, an additional C-terminal lysine residue is thought to be required for activity. This may be due to the presence of one hydrophilic residue between hydrophobic repeats. TIFF2025096309000099.tif5128 may be the cause.

[0257] Example 15 Derivatives of Peptides P15 / P20 that Elicit an HR Response in Tobacco To determine the minimum array required for HR and identify important residues, the HR test (described in Example 1) was performed with the variants of P15 / P20. The following peptides in Table 18 were determined to be positive for HR.

[0258] (Table 18) TIFF2025096309000100.tif64168

[0259] Example 16 Derivatives of Peptides P17 / P18 That Bring About an HR Reaction in Tobacco To determine the minimum array required for HR and identify important residues, the HR test (described in Example 1) was performed with the variants of P17 and P18. The following peptides in Table 19 were determined to be positive for HR.

[0260] (Table 19) TIFF2025096309000101.tif68166 * =N-terminal sequence TIFF2025096309000102.tif4128

[0261] Example 17 Derivatives of Peptide P19 That Bring About an HR Reaction in Tobacco To determine the minimum array required for HR and identify important residues, the HR test (described in Example 1) was performed with the variants of P19. The following peptides in Table 20 were determined to be positive for HR.

[0262] (Table 20) TIFF2025096309000103.tif57162

[0263] P19 and P19-1 show HR, but it is important to note that they do not completely match the consensus HR box sequence (SEQ ID NO:93). However, the addition of the context sequence in P19-2 and P19-3 results in a sequence that matches the consensus. Presumably, the additional isoleucine residues in P19, and P19-1 (N-terminal isoleucine and IGDN sequence) would increase the properties for HR induction.

[0264] Example 18 Induced resistance of tobacco against infection by tobacco mosaic virus In tobacco, peptides were tested for induction of resistance against tobacco mosaic virus (TMV). Briefly, three 6 - 8 week-old tobacco plants per group were selected (samples and controls). The bottom leaves of the plants were covered, and the plants were sprayed with a solution of water (negative control), peptide, or Proact (positive control). The spray was applied until the leaves were thoroughly wet and droplets of liquid were visible dripping from the leaves. The plants were then dried and the leaf covers were removed.

[0265] Then, three days after the treatment, diatomaceous earth was lightly sprinkled on the pre-covered leaves and the opposite leaves of the plants, and 20 μl of a 1.7 μg / ml purified tobacco mosaic virus solution was applied. The TMV solution was then spread over the entire leaf surface by gently rubbing the solution and diatomaceous earth over the entire leaf surface. Two minutes after inoculation, the diatomaceous earth was rinsed off the leaves with water. Three days after TMV inoculation, the leaves were scored based on the number of TMV lesions observed. The leaves were also scored for signs of hypersensitive reaction, including yellowing and withering of the infected leaves.

[0266] The efficacy described in Table 21 relates to the % reduction in TMV lesions in treated plants compared to UTC plants. The reduction of TMV in the covered leaves indicates a systemic immune response in the plants, while the reduction in the non-covered leaves indicates a local response. The asterisk indicates that the P-value derived from the t-test was <0.05.

[0267] (Table 21) Summary of TMV resistance TIFF2025096309000104.tif152168

[0268] Overall, the peptides that induced hypersensitive reactions in tobacco also conferred strong resistance to TMV. The peptides provided resistance in the treated leaves. However, the peptides also brought about "systemic acquired resistance," which triggers signaling in which an immune response in one part of the plant increases immunity in other parts of the plant. This was shown by a decrease in TMV infection in the coated leaves that were not directly treated with the peptide. Peptides that brought about particularly strong immune responses included a part of the minimal HR box peptide sequence: P14d, P25-11, and P30-3.

[0269] Example 19 Effect of Peptide Seed Treatment on Root and Shoot Growth Peptides were tested for their biological effects on the allocation of growth resources to shoots (above ground) and roots (below ground). Peptides were dissolved at 0.2, 2, or 5 μg / ml in a total volume of 100 ml of deionized water. Maize or soybean seeds were then immersed in the peptide solution for 1 hour. Untreated control (UTC) plants were immersed in deionized water. A clear plastic 300-ml beverage cup (Solo® (registered trademark), Dart Container Corporation) was prepared for planting by making a cross mark at the bottom and equally quartering the bottom. The cup was then filled with Sunshine Mix #1 soil (SunGro Horticulture) sifted to 1 / 4 inch. 100 ml of water was added to the soil. The treated seeds were then planted by gently pressing the seeds into the top of the soil. The seeds were then covered with an additional 50 ml of coarse soil. The seeds were germinated and grown for 12 - 14 days.

[0270] For each plant, the shoot length was measured as the distance from the soil to the lightly extended tip of the topmost leaf. Plants that did not germinate or showed stunted growth were excluded from the test. Stunted growth was defined as the cotyledons not being fully developed at the time of data collection or having developed true leaves with a visible area less than 1 / 2 of the average leaf area of the treatment group. Generally, 30 seeds were planted per treatment group, and 15 - 25 plants were used for data collection.

[0271] Root growth was estimated by counting the number of times the primary roots crossed the quarter marks at the bottom of the cup. These were often observed along the bottom perimeter of the cup, but some were visible along the side of the container and were counted similarly if they crossed the vertical extension of the quarter lines. This number was divided by 4 to generate a root growth index. This index was found to correlate approximately 90% with the total measured length of the primary roots (the sum of the lengths of all primary roots after rinsing the soil off the roots and measuring directly).

[0272] (Table 22) Summary of root and shoot growth TIFF2025096309000105.tif69138

[0273] Although the basic concepts of the present invention have been described as above, it will be apparent to those skilled in the art that the above detailed disclosure is intended merely as an example and not as a limitation. Even if not explicitly stated herein, various modifications, improvements, and variations will be recalled and intended by those skilled in the art. These modifications, improvements, and variations are intended to be suggested by this specification and are within the spirit and scope of the present invention. In addition, the order of the processing elements or arrays described, or the use of numbers, letters, or other symbols, is not intended to limit the processes described in the claims to any particular order, unless specified in the claims. Therefore, the present invention is limited only by the following claims and their equivalents.

[0274] Sequence information SEQUENCE LISTING <110> Plant Health Care, Inc. <120> HYPERSENSITIVE RESPONSE ELICITOR PEPTIDES AND USE THEREOF <150> US 62 / 058,535 <151> 2014-10-01 <150> US 62 / 140,789 <151> 2015-03-31 <160> 232 <170> PatentIn version 3.5 <210> 1 <211> 23 <212> PRT <213> Artificial <220> <223> P1 / P4 consensus <220> <221> MISC_FEATURE <222> (1)..(1) <223> X at position 1 is optional, S, N, D, isoD, G, A, or S <220> <221> MISC_FEATURE <222> (2)..(2) <223> X at position 2 is optional, Q, E, g-glutamate, G, A, or S <220> <221> MISC_FEATURE <222> (8)..(8) <223> X at position 8 is Q, E, g-glutamate, G, A, or S <220> <221> MISC_FEATURE <222> (9)..(9) <223> X at position 9 is L, I, F, or V <220> <221> MISC_FEATURE <222> (10)..(10) <223> X at position 10 is optional, D or isoD <220> <221> MISC_FEATURE <222> (11)..(11) <223> X at position 11 is Q, E, g-glutamate, G, A, or S <220> <221> MISC_FEATURE <222> (12)..(12) <223> X at position 12 is M, L, I, or F <220> <221> MISC_FEATURE <222> (13)..(13) <223> X at position 13 is M, L, I <220> <221> MISC_FEATURE <222> (14)..(14) <223> X at position 14 is optional, any hydrophilic amino acid, preferably C, S, T, A, D, isoD, K, or Q <220> <221> MISC_FEATURE <222> (15)..(15) <223> X at position 15 is Q, E, g-glutamate, G, A, S, K, or I <220> <221> MISC_FEATURE <222> (16)..(16) <223> X at position 16 is M, L, I, V, F <220> <221> MISC_FEATURE <222> (17)..(17) <223> X at position 17 is M, L, I, A, or V <220> <221> MISC_FEATURE <222> (18)..(18) <223> X at position 18 is Q, E, g-glutamate, G, A, S, M, T, or K <220> <221> MISC_FEATURE <222> (19)..(19) <223> X at position 19 is A, D, isoD, S, V, T, K, R, E, H, or G <220> <221> MISC_FEATURE <222> (20)..(20) <223> X at position 20 is M, L, or I <220> <221> MISC_FEATURE <222> (21)..(21) <223> X at position 21 is M, L, I, V, S, or F <220> <221> MISC_FEATURE <222> (22)..(22) <223> X at position 22 is Q, E, g-glutamate, G, A, or S <220> <221> MISC_FEATURE <222> (23)..(23) <223> The X at position 23 is P, Q, E, g-glutamate, G, A, or S <400> 1 Xaa Xaa Gly Ile Ser Glu Lys Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 1 5 10 15 Xaa Xaa Xaa Xaa Xaa Xaa Xaa 20 <210> 2 <211> 23 <212> PRT <213> Artificial <220> <223> P4 consensus <220> <221> MISC_FEATURE <222> (2)..(2) <223> The X at position 2 is Q, E, g-glutamate, G, A, or S <220> <221> MISC_FEATURE <222> (8)..(8) <223> The X at position 8 is Q, E, g-glutamate, G, A, or S <220> <221> MISC_FEATURE <222> (9)..(9) <223> The X at position 9 is L, A, D, isoD, I, F, or V <220> <221> MISC_FEATURE <222> (11)..(11) <223> The X at position 11 is Q, E, g-glutamate, G, A, or S <220> <221> MISC_FEATURE <222> (12)..(12) <223> X at position 12 is L, D, isoD, I, or F <220> <221> MISC_FEATURE <222> (13)..(13) <223> X at position 13 is L, I, V, or F <220> <221> MISC_FEATURE <222> (14)..(14) <223> X at position 14 is any hydrophilic amino acid, preferably C, S, or T <220> <221> MISC_FEATURE <222> (15)..(15) <223> X at position 15 is Q, E, g-glutamate, G, A, S, K, or I <220> <221> MISC_FEATURE <222> (16)..(16) <223> X at position 16 is L, A, I, V, M, or F <220> <221> MISC_FEATURE <222> (17)..(17) <223> X at position 17 is I, S, or F <220> <221> MISC_FEATURE <222> (18)..(18) <223> X at position 18 is Q, E, g-glutamate, G, A, S <220> <221> MISC_FEATURE <222> (20)..(20) <223> X at position 20 is L, I, V, F <220> <221> MISC_FEATURE <222> (21)..(21) <223> X at position 21 is L or F <220> <221> MISC_FEATURE <222> (22)..(22) <223> X at position 22 is Q, E, g-glutamate, G, A, or S <400> 2 Ser Xaa Gly Ile Ser Glu Lys Xaa Xaa Asp Xaa Xaa Xaa Xaa Xaa Xaa 1 5 10 15 Xaa Xaa Ala Xaa Xaa Xaa Pro 20 <210> 3 <211> 23 <212> PRT <213> Artificial <220> <223> P1 consensus <220> <221> MISC_FEATURE <222> (1)..(1) <223> X at position 1 is N, D, isoD, G, A, or S <220> <221> MISC_FEATURE <222> (2)..(2) <223> X at position 2 is Q, E, g-glutamate, G, A, or S <220> <221> MISC_FEATURE <222> (8)..(8) <223> The X at position 8 is Q, E, γ-glutamate, G, A, or S <220> <221> MISC_FEATURE <222> (11)..(11) <223> The X at position 11 is Q, E, γ-glutamate, G, A, or S <220> <221> MISC_FEATURE <222> (15)..(15) <223> The X at position 15 is Q, E, γ-glutamate, G, A, S <220> <221> MISC_FEATURE <222> (18)..(18) <223> The X at position 18 is M, T, K, E, γ-glutamate, G, A, or S <220> <221> MISC_FEATURE <222> (22)..(22) <223> The X at position 22 is Q, E, γ-glutamate, G, A, or S <220> <221> MISC_FEATURE <222> (23)..(23) <223> The X at position 23 is Q, E, γ-glutamate, G, A, or S <400> 3 Xaa Xaa Gly Ile Ser Glu Lys Xaa Leu Asp Xaa Leu Leu Thr Xaa Leu 1 5 10 15 Ile Xaa Ala Leu Leu Xaa Xaa 20 <210> 4 <211> 23 <212> PRT <213> Artificial <220> <223> P1 <400> 4 Asn Gln Gly Ile Ser Glu Lys Gln Leu Asp Gln Leu Leu Thr Gln Leu 1 5 10 15 Ile Met Ala Leu Leu Gln Gln 20 <210> 5 <211> 23 <212> PRT <213> Artificial <220> <223> P4 <400> 5 Ser Gln Gly Ile Ser Glu Lys Gln Leu Asp Gln Leu Leu Cys Gln Leu 1 5 10 15 Ile Gln Ala Leu Leu Gln Pro 20 <210> 6 <211> 23 <212> PRT <213> Artificial <220> <223> P4-14S <400> 6 Ser Gln Gly Ile Ser Glu Lys Gln Leu Asp Gln Leu Leu Ser Gln Leu 1 5 10 15 Ile Gln Ala Leu Leu Gln Pro 20 <210> 7 <211> 23 <212> PRT <213> Artificial <220> <223> P4-14S-18S <400> 7 Ser Gln Gly Ile Ser Glu Lys Gln Leu Asp Gln Leu Leu Ser Gln Leu 1 5 10 15 Ile Ser Ala Leu Leu Gln Pro 20 <210> 8 <211> 23 <212> PRT <213> Artificial <220> <223> P4-14S-2,18E <400> 8 Ser Glu Gly Ile Ser Glu Lys Gln Leu Asp Gln Leu Leu Ser Gln Leu 1 5 10 15 Ile Glu Ala Leu Leu Gln Pro 20 <210> 9 <211> 23 <212> PRT <213> Artificial <220> <223> P4-2E-8E <400> 9 Ser Glu Gly Ile Ser Glu Lys Glu Leu Asp Gln Leu Leu Ser Gln Leu 1 5 10 15 Ile Gln Ala Leu Leu Gln Pro 20 <210> 10 <211> 23 <212> PRT <213> Artificial <220> <223> P4-2E-8E-15E <400> 10 Ser Glu Gly Ile Ser Glu Lys Glu Leu Asp Gln Leu Leu Ser Glu Leu 1 5 10 15 Ile Gln Ala Leu Leu Gln Pro 20 <210> 11 <211> 23 <212> PRT <213> Artificial <220> <223> P4-allE <400> 11 Ser Glu Gly Ile Ser Glu Lys Glu Leu Asp Glu Leu Leu Ser Glu Leu 1 5 10 15 Ile Glu Ala Leu Leu Gln Pro 20 <210> 12 <211> 14 <212> PRT <213> Artificial <220> <223> P15 / 20 min consensus <220> <221> MISC_FEATURE <222> (7)..(7) <223> X of position 7 is optional, any amino acid <220> <221> MISC_FEATURE <222> (10)..(10) <223> The X at position 10 is M, E, γ-glutamate, G, A, S, T, or K <220> <221> MISC_FEATURE <222> (14)..(14) <223> The X at position 14 is optional, Q, E, γ-glutamate, G, A, S <400> 12 Ile Ala Lys Leu Ile Ser Xaa Leu Ile Xaa Ser Leu Leu Xaa 1 5 10 <210> 13 <211> 19 <212> PRT <213> Artificial <220> <223> P14d consensus <220> <221> MISC_FEATURE <222> (1)..(1) <223> The X at position 1 is Q, N, D, E, γ-glutamate, isoD, or S <220> <221> MISC_FEATURE <222> (2)..(2) <223> The X at position 2 can be D, E, γ-glutamate, isoD <220> <221> MISC_FEATURE <222> (3)..(3) <223> The X at position 3 can be P, D, E, isoD, or γ-glutamate <220> <221> MISC_FEATURE <222> (4)..(4) <223> X at position 4 can be M, A, S, D, E, isoD, or g-glutamate <220> <221> MISC_FEATURE <222> (5)..(5) <223> X at position 5 can be Q, E, or g-glutamate <220> <221> MISC_FEATURE <222> (6)..(6) <223> X at position 6 can be A, E, or g-glutamate <220> <221> MISC_FEATURE <222> (8)..(8) <223> X at position 8 can be M, L, E, Q, D, N, G, A, S, isoD, or g-glutamate <220> <221> MISC_FEATURE <222> (9)..(9) <223> X at position 9 can be Q, N, E, D, G, A, S, isoD or, g-glutamate <220> <221> MISC_FEATURE <222> (12)..(12) <223> X at position 12 can be Q, N, E, D, G, A, S, isoD or, g-glutamate <220> <221> MISC_FEATURE <222> (13)..(13) <223> X at position 13 can be Q, N, E, D, G, A, S, isoD or, g-glutamate <220> <221> MISC_FEATURE <222> (16)..(16) <223> X at position 16 can be K, Q, N, E, D, R, G, A, or S <400> 13 Xaa Xaa Xaa Xaa Xaa Xaa Leu Xaa Xaa Leu Leu Xaa Xaa Leu Val Xaa 1 5 10 15 Leu Leu Lys <210> 14 <211> 13 <212> PRT <213> Artificial <220> <223> P14d min consensus <220> <221> MISC_FEATURE <222> (2)..(2) <223> X at position 2 can be M, L, E, Q, D, N, G, A, S, isoD, or g-glutamate <220> <221> MISC_FEATURE <222> (3)..(3) <223> X at position 3 can be Q, N, E, D, G, A, S, isoD or, g-glutamate <220> <221> MISC_FEATURE <222> (6)..(6) <223> X at position 6 is Q, N, E, D, G, A, S, isoD or, g-glutamate <220> <221> MISC_FEATURE <222> (7)..(7) <223> X at position 7 can be Q, N, E, D, G, A, S, isoD or, g-glutamate <220> <221> MISC_FEATURE <222> (10)..(10) <223> X at position 10 can be K, Q, N, E, D, R, G, A, or S <400> 14 Leu Xaa Xaa Leu Leu Xaa Xaa Leu Val Xaa Leu Leu Lys 1 5 10 <210> 15 <211> 14 <212> PRT <213> Artificial <220> <223> P3min consensus <220> <221> MISC_FEATURE <222> (1)..(1) <223> X at position 1 is L or M <220> <221> MISC_FEATURE <222> (2)..(2) <223> X at position 2 can be Q, N, E, g-glutamate, D, isoD, T, S, A, or G <220> <221> MISC_FEATURE <222> (3)..(3) <223> X at position 3 can be Q, N, E, g-glutamate, D, isoD, T, S, A, or G <220> <221> MISC_FEATURE <222> (6)..(6) <223> X at position 6 can be K, Q, N, E, g-glutamate, D, isoD, T, S, A, or G <220> <221> MISC_FEATURE <222> (7)..(7) <223> X at position 7 is L or M <220> <221> MISC_FEATURE <222> (9)..(9) <223> X at position 9 can be E, g-glutamate, D, isoD, Q, N, T, S, A, or G <220> <221> MISC_FEATURE <222> (10)..(10) <223> X at position 10 can be A, G, S, T, E, g-glutamate, D, isoD, Q, or N <220> <221> MISC_FEATURE <222> (12)..(12) <223> X at position 12 is L or M <220> <221> MISC_FEATURE <222> (13)..(13) <223> X at position 13 can be Q, N, E, g-glutamate, D, isoD, T, S, A, or G <220> <221> MISC_FEATURE <222> (14)..(14) <223> X at position 14 can be Q, N, E, g-glutamate, D, isoD, T, S, A, or G <400> 15 Xaa Xaa Xaa Leu Leu Xaa Xaa Phe Xaa Xaa Ile Xaa Xaa Xaa 1 5 10 <210> 16 <211> 12 <212> PRT <213> Artificial <220> <223> P25 consensus <220> <221> MISC_FEATURE <222> (2)..(2) <223> X at position 2 can be Q, N, E, g-glutamate, D, isoD, T, S, A, or G <220> <221> MISC_FEATURE <222> (3)..(3) <223> X at position 3 can be K, Q, N, E, g-glutamate, D, isoD, T, S, A, or G <220> <221> MISC_FEATURE <222> (5)..(5) <223> X at position 5 can be L or M <220> <221> MISC_FEATURE <222> (6)..(6) <223> X at position 6 can be K, Q, N, E, g-glutamate, D, isoD, T, S, A, or G <220> <221> MISC_FEATURE <222> (9)..(9) <223> X at position 9 can be E, g-glutamate, D, isoD, Q, N, T, S, A, or G <220> <221> MISC_FEATURE <222> (10)..(10) <223> X at position 10 can be A, G, S, T, E, g-glutamate, D, isoD, Q, or N <400> 16 Leu Xaa Xaa Leu Xaa Xaa Ile Leu Xaa Xaa Leu Val 1 5 10 <210> 17 <211> 16 <212> PRT <213> Artificial <220> <223> P25 consensus <220> <221> MISC_FEATURE <222> (2)..(2) <223> X at position 2 can be T, S, A, G, D, isoD, E, g-glutamate, Q, or N <220> <221> MISC_FEATURE <222> (3)..(3) <223> X at position 3 can be G, T, S, A, D, isoD, E, g-glutamate, Q, or N <220> <221> MISC_FEATURE <222> (6)..(6) <223> X at position 6 can be Q, N, E, g-glutamate, D, isoD, T, S, A, or G <220> <221> MISC_FEATURE <222> (7)..(7) <223> X at position 7 can be K, Q, N, E, g-glutamate, D, isoD, T, S, A, or G <220> <221> MISC_FEATURE <222> (9)..(9) <223> X at position 9 can be L or M <220> <221> MISC_FEATURE <222> (10)..(10) <223> X at position 10 can be K, Q, N, E, g-glutamate, D, isoD, T, S, A, or G <220> <221> MISC_FEATURE <222> (13)..(13) <223> X at position 13 can be E, g-glutamate, D, isoD, Q, N, T, S, A, or G <220> <221> MISC_FEATURE <222> (14)..(14) <223> X at position 14 can be A, G, S, T, E, g-glutamate, D, isoD, Q, or N <220> <221> MISC_FEATURE <222> (16)..(16) <223> V at position 16 is optional <400> 17 Leu Xaa Xaa Val Leu Xaa Xaa Leu Xaa Xaa Ile Leu Xaa Xaa Leu Val 1 5 10 15 <210> 18 <211> 27 <212> PRT <213> Artificial <220> <223> P17 / 18 <220> <221> MISC_FEATURE <222> (1)..(1) <223> X at position 1 can be any amino acid, but preferably Q, S, E, g-glutamate, A, T, G, D, isoD, N, K, or R <220> <221> MISC_FEATURE <222> (2)..(2) <223> X at position 2 can be any amino acid, but preferably Q, S, E, g-glutamate, A, T, G, D, isoD, N, K, or R <220> <221> MISC_FEATURE <222> (3)..(3) <223> The X at position 3 can be any amino acid, but is preferably P, Q, S, E, g-glutamate, A, T, G, D, isoD, N, K, or R <220> <221> MISC_FEATURE <222> (4)..(4) <223> The X at position 4 can be any amino acid, but is preferably I, Q, S, E, g-glutamate, A, T, G, D, N, isoD, K, or R <220> <221> MISC_FEATURE <222> (5)..(5) <223> The X at position 5 can be any amino acid, but is preferably D, isoD, S, E, g-glutamate, A, T, G, N, Q, K, or R <220> <221> MISC_FEATURE <222> (6)..(6) <223> The X at position 6 can be any amino acid, but is preferably R, Q, S, E, g-glutamate, A, T, G, D, isoD, N, or K <220> <221> MISC_FEATURE <222> (7)..(7) <223> The amino acid at position 7 can be any amino acid, but is preferably Q, S, E, g-glutamate, A, T, G, D, isoD, N, K, or R <220> <221> MISC_FEATURE <222> (8)..(8) <223> The amino acid at position 8 can be any amino acid, but is preferably T, Q, S, E, g-glutamate, A, G, D, isoD, N, K, or R <220> <221> MISC_FEATURE <222> (9)..(9) <223> The amino acid at position 9 can be any amino acid, but is preferably I, Q, S, E, g-glutamate, A, T, G, D, isoD, N, K, or R <220> <221> MISC_FEATURE <222> (10)..(10) <223> The amino acid at position 10 can be any amino acid, but is preferably E, g-glutamate, Q, S, A, T, G, D, isoD, N, K, or R <220> <221> MISC_FEATURE <222> (11)..(11) <223> The amino acid at position 11 can be any amino acid, but is preferably Q, S, E, g-glutamate, A, T, G, D, isoD, N, K, or R <220> <221> MISC_FEATURE <222> (12)..(12) <223> X at position 12 can be L or M <220> <221> MISC_FEATURE <222> (13)..(13) <223> X at position 13 can be any amino acid, but preferably A, S, T, G, D, isoD, E, g-glutamate, Q, N, K, or R <220> <221> MISC_FEATURE <222> (14)..(14) <223> X at position 14 can be any amino acid, but preferably Q, A, S, T, G, D, isoD, E, g-glutamate, N, K, or R <220> <221> MISC_FEATURE <222> (17)..(17) <223> X at position 17 can be any amino acid, but preferably A, S, T, G, D, isoD, E, g-glutamate, Q, N, K, or R <220> <221> MISC_FEATURE <222> (18)..(18) <223> X at position 18 can be any amino acid, but preferably Q, A, S, T, G, D, isoD, E, g-glutamate, N, K, or R <220> <221> MISC_FEATURE <222> (21)..(21) <223> The X at position 21 can be any amino acid, but preferably K, A, S, T, G, D, isoD, E, g-glutamate, Q, N, or R <220> <221> MISC_FEATURE <222> (22)..(22) <223> The X at position 22 can be any amino acid, but preferably S, A, T, G, D, isoD, E, g-glutamate, Q, N, K, or R <220> <221> MISC_FEATURE <222> (25)..(25) <223> The X at position 25 can be any amino acid, but preferably S, A, T, G, D, isoD, E, g-glutamate, Q, N, K, or R <220> <221> MISC_FEATURE <222> (26)..(26) <223> The X at position 26 can be any amino acid, but preferably P, S, A, T, G, D, isoD, E, g-glutamate, Q, N, K, or R <220> <221> MISC_FEATURE <222> (27)..(27) <223> The X at position 27 can be any amino acid, but preferably Q, S, A, T, G, D, isoD, E, g-glutamate, N, K, or R <400> 18 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Leu Leu 1 5 10 15 Xaa Xaa Leu Leu Xaa Xaa Leu Leu Xaa Xaa Xaa 20 25 <210> 19 <211> 23 <212> PRT <213> Artificial <220> <223> P4-14s-9v <400> 19 Ser Gln Gly Ile Ser Glu Lys Gln Val Asp Gln Leu Leu Ser Gln Leu 1 5 10 15 Ile Gln Ala Leu Leu Gln Pro 20 <210> 20 <211> 23 <212> PRT <213> Artificial <220> <223> P4-14s-9F <400> 20 Ser Gln Gly Ile Ser Glu Lys Gln Phe Asp Gln Leu Leu Ser Gln Leu 1 5 10 15 Ile Gln Ala Leu Leu Gln Pro 20 <210> 21 <211> 27 <212> PRT <213> Artificial <220> <223> P17 / 18 <220> <221> MISC_FEATURE <222> (1)..(1) <223> The X at position 1 can be any amino acid, but preferably Q, S, E, g-glutamate, A, T, G, D, isoD, N, K, or R <220> <221> MISC_FEATURE <222> (2)..(2) <223> The X at position 2 can be any amino acid, but preferably Q, S, E, g-glutamate, A, T, G, D, isoD, N, K, or R <220> <221> MISC_FEATURE <222> (3)..(3) <223> The X at position 3 can be any amino acid, but preferably P, Q, S, E, g-glutamate, A, T, G, D, isoD, N, K, or R <220> <221> MISC_FEATURE <222> (4)..(4) <223> The X at position 4 can be any amino acid, but preferably I, Q, S, E, g-glutamate, A, T, G, D, N, isoD, K, or R <220> <221> MISC_FEATURE <222> (5)..(5) <223> The amino acid at position 5 can be any amino acid, but is preferably D, isoD, S, E, g-glutamate, A, T, G, N, Q, K, or R <220> <221> MISC_FEATURE <222> (6)..(6) <223> The amino acid at position 6 can be any amino acid, but is preferably R, Q, S, E, g-glutamate, A, T, G, D, isoD, N, or K <220> <221> MISC_FEATURE <222> (7)..(7) <223> The amino acid at position 7 can be any amino acid, but is preferably Q, S, E, g-glutamate, A, T, G, D, isoD, N, K, or R <220> <221> MISC_FEATURE <222> (8)..(8) <223> The amino acid at position 8 can be any amino acid, but is preferably T, Q, S, E, g-glutamate, A, G, D, isoD, N, K, or R <220> <221> MISC_FEATURE <222> (9)..(9) <223> The amino acid at position 9 can be any amino acid, but is preferably I, Q, S, E, g-glutamate, A, T, G, D, isoD, N, K, or R <220> <221> MISC_FEATURE <222> (10)..(10) <223> The X at position 10 can be any amino acid, but preferably E, g-glutamate, Q, S, A, T, G, D, isoD, N, K, or R <220> <221> MISC_FEATURE <222> (11)..(11) <223> The X at position 11 can be any amino acid, but preferably Q, S, E, g-glutamate, A, T, G, D, isoD, N, K, or R <220> <221> MISC_FEATURE <222> (12)..(12) <223> The X at position 12 can be L or M <220> <221> MISC_FEATURE <222> (13)..(13) <223> The X at position 13 can be any amino acid, but preferably A, S, T, G, D, isoD, E, g-glutamate, Q, N, K, or R <220> <221> MISC_FEATURE <222> (14)..(14) <223> The X at position 14 can be any amino acid, but preferably Q, A, S, T, G, D, isoD, E, g-glutamate, N, K, or R <220> <221> MISC_FEATURE <222> (17)..(17) <223> The X at position 17 can be any amino acid, but is preferably A, S, T, G, D, isoD, E, g-glutamate, Q, N, K, or R <220> <221> MISC_FEATURE <222> (18)..(18) <223> The X at position 18 can be any amino acid, but is preferably Q, A, S, T, G, D, isoD, E, g-glutamate, N, K, or R <220> <221> MISC_FEATURE <222> (21)..(21) <223> The X at position 21 can be any amino acid, but is preferably K, A, S, T, G, D, isoD, E, g-glutamate, Q, N, or R <220> <221> MISC_FEATURE <222> (22)..(22) <223> The X at position 22 can be any amino acid, but is preferably S, A, T, G, D, isoD, E, g-glutamate, Q, N, K, or R <220> <221> MISC_FEATURE <222> (25)..(25) <223> The X at position 25 can be any amino acid, but is preferably S, A, T, G, D, isoD, E, g-glutamate, Q, N, K, or R <220> <221> MISC_FEATURE <222> (26)..(26) <223> X at position 26 can be any amino acid, but preferably P, S, A, T, G, D, isoD, E, g-glutamate, Q, N, K, or R <220> <221> MISC_FEATURE <222> (27)..(27) <223> X at position 27 can be any amino acid, but preferably Q, S, A, T, G, D, isoD, E, g-glutamate, N, K, or R <400> 21 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Leu Leu 1 5 10 15 Xaa Xaa Leu Leu Xaa Xaa Leu Leu Xaa Xaa Xaa 20 25 <210> 22 <211> 23 <212> PRT <213> Artificial <220> <223> P4-14s-12i <400> 22 Ser Gln Gly Ile Ser Glu Lys Gln Leu Asp Gln Ile Leu Ser Gln Leu 1 5 10 15 Ile Gln Ala Leu Leu Gln Pro 20 <210> 23 <211> 23 <212> PRT <213> Artificial <220> <223> P4-14s-12f <400> 23 Ser Gln Gly Ile Ser Glu Lys Gln Leu Asp Gln Phe Leu Ser Gln Leu 1 5 10 15 Ile Gln Ala Leu Leu Gln Pro 20 <210> 24 <211> 23 <212> PRT <213> Artificial <220> <223> P4-14s-13i <400> 24 Ser Gln Gly Ile Ser Glu Lys Gln Leu Asp Gln Leu Ile Ser Gln Leu 1 5 10 15 Ile Gln Ala Leu Leu Gln Pro 20 <210> 25 <211> 13 <212> PRT <213> Artificial <220> <223> P17 / 18 min consensus <220> <221> MISC_FEATURE <222> (1)..(1) <223> X at position 1 can be L or M <220> <221> MISC_FEATURE <222> (2)..(2) <223> The X at position 2 can be any amino acid, but is preferably A, S, T, G, D, isoD, E, g-glutamate, Q, N, K, or R <220> <221> MISC_FEATURE <222> (3)..(3) <223> The X at position 3 can be any amino acid, but is preferably Q, A, S, T, G, D, isoD, E, g-glutamate, N, K, or R <220> <221> MISC_FEATURE <222> (6)..(6) <223> The X at position 6 can be any amino acid, but is preferably A, S, T, G, D, isoD, E, g-glutamate, Q, N, K, or R <220> <221> MISC_FEATURE <222> (7)..(7) <223> The X at position 7 can be any amino acid, but is preferably Q, A, S, T, G, D, isoD, E, g-glutamate, N, K, or R <220> <221> MISC_FEATURE <222> (10)..(10) <223> The X at position 10 can be any amino acid, but is preferably K, A, S, T, G, D, isoD, E, g-glutamate, Q, N, or R <220> <221> MISC_FEATURE <222> (11)..(11) <223> X at position 11 can be any amino acid, but preferably S, A,T, G, D, isoD, E, g-glutamate, Q, N, K, or R <400> 25 Xaa Xaa Xaa Leu Leu Xaa Xaa Leu Leu Xaa Xaa Leu Leu 1 5 10 <210> 26 <211> 13 <212> PRT <213> Artificial <220> <223> P19 consensus <220> <221> MISC_FEATURE <222> (1)..(1) <223> X at position 1 is optional and can be L, I, V, F, or M <220> <221> MISC_FEATURE <222> (3)..(3) <223> X at position 3 can be any amino acid, but preferably K, A, S, T, G, D, isoD, E, g-glutamate, Q, N, or R <220> <221> MISC_FEATURE <222> (4)..(4) <223> X at position 4 can be any amino acid, but preferably A, S, T, G, D, isoD, E, g-glutamate, Q, N, K, or R <220> <221> MISC_FEATURE <222> (5)..(5) <223> X at position 5 can be L or M <220> <221> MISC_FEATURE <222> (7)..(7) <223> X at position 7 can be any amino acid, but preferably K, A, S, T, G, D, isoD, E, g-glutamate, Q, N, or R <220> <221> MISC_FEATURE <222> (10)..(10) <223> X at position 10 can be any amino acid, but preferably A, S, T, G, D, isoD, E, g-glutamate, Q, N, K, or R <220> <221> MISC_FEATURE <222> (11)..(11) <223> X at position 11 can be any amino acid, but preferably R, A, S, T, G, D, isoD, E, g-glutamate, Q, N, or K <220> <221> MISC_FEATURE <222> (12)..(12) <223> X at position 12 can be L, I, V, F, or M <220> <221> MISC_FEATURE <222> (13)..(13) <223> X at position 13 can be L, I, V, F, or M <400> 26 Xaa Leu Xaa Xaa Xaa Leu Xaa Leu Ile Xaa Xaa Xaa Xaa 1 5 10 <210> 27 <211> 23 <212> PRT <213> Artificial <220> <223> P4-14s-15a <400> 27 Ser Gln Gly Ile Ser Glu Lys Gln Leu Asp Gln Leu Leu Ser Ala Leu 1 5 10 15 Ile Gln Ala Leu Leu Gln Pro 20 <210> 28 <211> 23 <212> PRT <213> Artificial <220> <223> P4-14s-15k <400> 28 Ser Gln Gly Ile Ser Glu Lys Gln Leu Asp Gln Leu Leu Ser Lys Leu 1 5 10 15 Ile Gln Ala Leu Leu Gln Pro 20 <210> 29 <211> 23 <212> PRT <213> Artificial <220> <223> P4-14s-15s <400> 29 Ser Gln Gly Ile Ser Glu Lys Gln Leu Asp Gln Leu Leu Ser Ser Leu 1 5 10 15 Ile Gln Ala Leu Leu Gln Pro 20 <210> 30 <211> 23 <212> PRT <213> Artificial <220> <223> P4-14s-15i <400> 30 Ser Gln Gly Ile Ser Glu Lys Gln Leu Asp Gln Leu Leu Ser Ile Leu 1 5 10 15 Ile Gln Ala Leu Leu Gln Pro 20 <210> 31 <211> 20 <212> PRT <213> Artificial <220> <223> polyE-min2p4 <400> 31 Ser Glu Glu Glu Glu Glu Leu Asp Gln Leu Leu Ser Gln Leu Ile Gln 1 5 10 15 Ala Leu Leu Gln 20 <210> 32 <211> 23 <212> PRT <213> Artificial <220> <223> P4-14s-16i <400> 32 Ser Gln Gly Ile Ser Glu Lys Gln Leu Asp Gln Leu Leu Ser Gln Ile 1 5 10 15 Ile Gln Ala Leu Leu Gln Pro 20 <210> 33 <211> 19 <212> PRT <213> Artificial <220> <223> polyE-min3P4 <400> 33 Ser Glu Glu Glu Glu Glu Leu Asp Gln Leu Leu Ser Gln Leu Ile Gln 1 5 10 15 Ala Leu Leu <210> 34 <211> 23 <212> PRT <213> Artificial <220> <223> P4-14s-16f <400> 34 Ser Gln Gly Ile Ser Glu Lys Gln Leu Asp Gln Leu Leu Ser Gln Phe 1 5 10 15 Ile Gln Ala Leu Leu Gln Pro 20 <210> 35 <211> 22 <212> PRT <213> Artificial <220> <223> P4-NpolyR <400> 35 Arg Arg Arg Arg Arg Gly Gly Leu Asp Gln Leu Leu Ser Gln Leu Ile 1 5 10 15 Gln Ala Leu Leu Gln Pro 20 <210> 36 <211> 22 <212> PRT <213> Artificial <220> <223> P4-CpolyR <400> 36 Leu Asp Gln Leu Leu Ser Gln Leu Ile Gln Ala Leu Leu Gln Pro Gly 1 5 10 15 Gly Arg Arg Arg Arg Arg 20 <210> 37 <211> 23 <212> PRT <213> Artificial <220> <223> P4-14s-20i <400> 37 Ser Gln Gly Ile Ser Glu Lys Gln Leu Asp Gln Leu Leu Ser Gln Leu 1 5 10 15 Ile Gln Ala Ile Leu Gln Pro 20 <210> 38 <211> 22 <212> PRT <213> Artificial <220> <223> P4-NpolyK <400> 38 Lys Lys Lys Lys Lys Gly Gly Leu Asp Gln Leu Leu Ser Gln Leu Ile 1 5 10 15 Gln Ala Leu Leu Gln Pro 20 <210> 39 <211> 22 <212> PRT <213> Artificial <220> <223> P4-CpolyK <400> 39 Leu Asp Gln Leu Leu Ser Gln Leu Ile Gln Ala Leu Leu Gln Pro Gly 1 5 10 15 Gly Lys Lys Lys Lys Lys 20 <210> 40 <211> 23 <212> PRT <213> Artificial <220> <223> P4-14s-21f <400> 40 Ser Gln Gly Ile Ser Glu Lys Gln Leu Asp Gln Leu Leu Ser Gln Leu 1 5 10 15 Ile Gln Ala Leu Phe Gln Pro 20 <210> 41 <211> 23 <212> PRT <213> Artificial <220> <223> P1-2E <400> 41 Asn Glu Gly Ile Ser Glu Lys Gln Leu Asp Gln Leu Leu Thr Gln Leu 1 5 10 15 Ile Met Ala Leu Leu Gln Gln 20 <210> 42 <211> 23 <212> PRT <213> Artificial <220> <223> P1-18T <400> 42 Asn Gln Gly Ile Ser Glu Lys Gln Leu Asp Gln Leu Leu Thr Gln Leu 1 5 10 15 Ile Thr Ala Leu Leu Gln Gln 20 <210> 43 <211> 23 <212> PRT <213> Artificial <220> <223> P1-18E <400> 43 Asn Gln Gly Ile Ser Glu Lys Gln Leu Asp Gln Leu Leu Thr Gln Leu 1 5 10 15 Ile Glu Ala Leu Leu Gln Gln 20 <210> 44 <211> 23 <212> PRT <213> Artificial <220> <223> P1-18A <400> 44 Asn Gln Gly Ile Ser Glu Lys Gln Leu Asp Gln Leu Leu Thr Gln Leu 1 5 10 15 Ile Ala Ala Leu Leu Gln Gln 20 <210> 45 <211> 23 <212> PRT <213> Artificial <220> <223> P1-18K <400> 45 Asn Gln Gly Ile Ser Glu Lys Gln Leu Asp Gln Leu Leu Thr Gln Leu 1 5 10 15 Ile Lys Ala Leu Leu Gln Gln 20 <210> 46 <211> 23 <212> PRT <213> Artificial <220> <223> P1-2E,8E,11E,15E,18E <400> 46 Asn Glu Gly Ile Ser Glu Lys Glu Leu Asp Glu Leu Leu Thr Glu Leu 1 5 10 15 Ile Glu Ala Leu Leu Gln Gln 20 <210> 47 <211> 22 <212> PRT <213> Artificial <220> <223> P15b / P20 consensus <220> <221> MISC_FEATURE <222> (3)..(3) <223> X at position 3 is N, D, or isoD <220> <221> MISC_FEATURE <222> (6)..(6) <223> X at position 6 is Q, E, g-glutamate, G, A, or S <220> <221> MISC_FEATURE <222> (8)..(8) <223> X at position 8 is N, D, or isoD <220> <221> MISC_FEATURE <222> (15)..(15) <223> X at position 15 is optional and can be any amino acid <220> <221> MISC_FEATURE <222> (18)..(18) <223> X at position 18 is M, E, g-glutamate, G, A, S, T, or K <220> <221> MISC_FEATURE <222> (22)..(22) <223> X at position 22 is optional and can be Q, E, g-glutamate, G, A, or S <400> 47 Lys Pro Xaa Asp Ser Xaa Ser Xaa Ile Ala Lys Leu Ile Ser Xaa Leu 1 5 10 15 Ile Xaa Ser Leu Leu Xaa 20 <210> 48 <211> 45 <212> PRT <213> Artificial <220> <223> Wildtype P15 <400> 48 Gln Lys Asp Val Asn Phe Gly Thr Pro Asp Ser Thr Val Gln Asn Pro 1 5 10 15 Gln Asp Ala Ser Lys Pro Asn Asp Ser Gln Ser Asn Ile Ala Lys Leu 20 25 30 Ile Ser Ala Leu Ile Met Ser Leu Leu Gln Met Leu Thr 35 40 45 <210> 49 <211> 22 <212> PRT <213> Artificial <220> <223> P15b <400> 49 Lys Pro Asn Asp Ser Gln Ser Asn Ile Ala Lys Leu Ile Ser Ala Leu 1 5 10 15 Ile Met Ser Leu Leu Gln 20 <210> 50 <211> 22 <212> PRT <213> Artificial <220> <223> P15b-8D-18E <400> 50 Lys Pro Asn Asp Ser Gln Ser Asp Ile Ala Lys Leu Ile Ser Ala Leu 1 5 10 15 Ile Glu Ser Leu Leu Gln 20 <210> 51 <211> 22 <212> PRT <213> Artificial <220> <223> P15b-8D-18A <400> 51 Lys Pro Asn Asp Ser Gln Ser Asp Ile Ala Lys Leu Ile Ser Ala Leu 1 5 10 15 Ile Ala Ser Leu Leu Gln 20 <210> 52 <211> 22 <212> PRT <213> Artificial <220> <223> P15b-8D-18S <400> 52 Lys Pro Asn Asp Ser Gln Ser Asp Ile Ala Lys Leu Ile Ser Ala Leu 1 5 10 15 Ile Ser Ser Leu Leu Gln 20 <210> 53 <211> 22 <212> PRT <213> Artificial <220> <223> P15b-8D-18T <400> 53 Lys Pro Asn Asp Ser Gln Ser Asp Ile Ala Lys Leu Ile Ser Ala Leu 1 5 10 15 Ile Thr Ser Leu Leu Gln 20 <210> 54 <211> 22 <212> PRT <213> Artificial <220> <223> P15b-8D-18K <400> 54 Lys Pro Asn Asp Ser Gln Ser Asp Ile Ala Lys Leu Ile Ser Ala Leu 1 5 10 15 Ile Lys Ser Leu Leu Gln 20 <210> 55 <211> 22 <212> PRT <213> Artificial <220> <223> P15b-8D-6,18E <400> 55 Lys Pro Asn Asp Ser Glu Ser Asp Ile Ala Lys Leu Ile Ser Ala Leu 1 5 10 15 Ile Glu Ser Leu Leu Gln 20 <210> 56 <211> 22 <212> PRT <213> Artificial <220> <223> P15b-3,8D <400> 56 Lys Pro Asp Asp Ser Gln Ser Asp Ile Ala Lys Leu Ile Ser Ala Leu 1 5 10 15 Ile Met Ser Leu Leu Gln 20 <210> 57 <211> 22 <212> PRT <213> Artificial <220> <223> P15b-3,8D-6E <400> 57 Lys Pro Asp Asp Ser Glu Ser Asp Ile Ala Lys Leu Ile Ser Ala Leu 1 5 10 15 Ile Met Ser Leu Leu Gln 20 <210> 58 <211> 22 <212> PRT <213> Artificial <220> <223> P15b-3,8D-18E <400> 58 Lys Pro Asp Asp Ser Gln Ser Asp Ile Ala Lys Leu Ile Ser Ala Leu 1 5 10 15 Ile Glu Ser Leu Leu Gln 20 <210> 59 <211> 22 <212> PRT <213> Artificial <220> <223> P15b-3,8D-6,18E <400> 59 Lys Pro Asp Asp Ser Glu Ser Asp Ile Ala Lys Leu Ile Ser Ala Leu 1 5 10 15 Ile Glu Ser Leu Leu Gln 20 <210> 60 <211> 22 <212> PRT <213> Artificial <220> <223> P15b-3,8D-allE <400> 60 Lys Pro Asp Asp Ser Glu Ser Asp Ile Ala Lys Leu Ile Ser Ala Leu 1 5 10 15 Ile Glu Ser Leu Leu Glu 20 <210> 61 <211> 22 <212> PRT <213> Artificial <220> <223> P15b-8D-allE <400> 61 Lys Pro Asn Asp Ser Glu Ser Asp Ile Ala Lys Leu Ile Ser Ala Leu 1 5 10 15 Ile Glu Ser Leu Leu Glu 20 <210> 62 <211> 22 <212> PRT <213> Artificial <220> <223> P15b-3D-allE <400> 62 Lys Pro Asp Asp Ser Glu Ser Asn Ile Ala Lys Leu Ile Ser Ala Leu 1 5 10 15 Ile Glu Ser Leu Leu Glu 20 <210> 63 <211> 39 <212> PRT <213> Artificial <220> <223> P15a <400> 63 Asn Phe Gly Thr Pro Asp Ser Thr Val Gln Asn Pro Gln Asp Ala Ser 1 5 10 15 Lys Pro Asn Asp Ser Gln Ser Asn Ile Ala Lys Leu Ile Ser Ala Leu 20 25 30 Ile Met Ser Leu Leu Gln Met 35 <210> 64 <211> 23 <212> PRT <213> Artificial <220> <223> P15 <400> 64 Lys Pro Asn Asp Ser Gln Ser Asn Ile Ala Lys Leu Ile Ser Ala Leu 1 5 10 15 Ile Met Ser Leu Leu Gln Met 20 <210> 65 <211> 34 <212> PRT <213> Artificial <220> <223> P20 <400> 65 Gly Thr Pro Asp Ser Thr Val Gln Asn Pro Gln Asp Ala Ser Lys Pro 1 5 10 15 Asn Asp Ser Gln Ser Asn Ile Ala Lys Leu Ile Ser Leu Ile Met Ser 20 25 30 Leu Leu <210> 66 <211> 20 <212> PRT <213> Artificial <220> <223> P6 / 6a consensus <220> <221> MISC_FEATURE <222> (4)..(4) <223> X at position 4 is F or Y <220> <221> MISC_FEATURE <222> (6)..(6) <223> X at position 6 is Q, E, g-glutamate, G, A, or S <220> <221> MISC_FEATURE <222> (7)..(7) <223> X at position 7 is optional and can be M, E, g-glutamate, G, A, S, T, or K or L <220> <221> MISC_FEATURE <222> (9)..(9) <223> X at position 9 is M, E, g-glutamate, G, A, S, T, or K <220> <221> MISC_FEATURE <222> (10)..(10) <223> X at position 10 is H or N <220> <221> MISC_FEATURE <222> (14)..(14) <223> X at position 14 is E, g-glutamate, D, or isoD <...

Claims

1. 1. An isolated peptide consisting of the amino acid sequence of XXXXXXXXXXXX(L / M)XXLLXXLLXXLLXXX (SEQ ID NO:18, P17 / 18), optionally containing up to three additional amino acids at its C-terminus, Where: X at position 1 is optional and, if present, is Q; X at position 2 is optional and, if present, is Q; X at position 3 is optional and, when present, is P or S; X at position 4 is optional and, if present, is I, S, or E; X at position 5 is optional and, if present, is D; X at position 6 is optional and, when present, is R, S, E, D, or N; X at position 7 is optional and, if present, is Q, E, or D; X at position 8 is optional and, if present, is T, Q, S, or E; X at position 9 is optional and, if present, is I, Q, S, or E; X at position 10 is optional and, if present, is E, S, or K; X at position 11 is optional and, if present, is Q, E, or D; X at position 13 is A, D, or E; X at position 14 is Q, E, or K; X at position 17 is A, S, or E; X at position 18 is Q, A, or E; X at position 21 is K, E, or Q; X at position 22 is S, A, or E; X at position 25 is optional and, if present, is S, A, or E; X at position 26 is optional and, if present, is P or E; and X at position 27 is optional and, if present, is Q or E; When introduced into mechanically wounded plant leaf tissue, it induces a hypersensitive response. The isolated peptide.

2. 2. The isolated peptide of claim 1, which is (i) more stable than the polypeptide of SEQ ID NO: 162 when dissolved in water or an aqueous solution, or (ii) more resistant to chemical degradation than the polypeptide of SEQ ID NO: 162 when dissolved in an aqueous buffer containing a biocide.

3. 2. The isolated peptide of claim 1, comprising one of the amino acid sequences of SEQ ID NOs: 83-88, 119, 163-167, 228, 229, and 231.

4. 2. The isolated peptide of claim 1, comprising one of the amino acid sequences of SEQ ID NOs: 83-88, 119, 163-167, 228, 229, and 231.

5. 2. The isolated peptide of claim 1, wherein one or more of the amino acids at positions 1 to 11 and / or the amino acids at positions 25 to 27 are absent.

6. 2. The isolated peptide of claim 1, which is free of lysine.

7. 2. The isolated peptide of claim 1, having a stability of at least 66% in water or an aqueous solution at 50° C. for 7 days.

8. 2. The isolated peptide of claim 1, which is at least 90% pure.

9. 8. The isolated peptide of any one of claims 1 to 7, which is a fusion polypeptide comprising a second amino acid sequence coupled to said amino acid sequence via a peptide bond.

10. 10. The isolated peptide of claim 9, wherein the second amino acid sequence comprises a purification tag.

11. 11. The isolated peptide of claim 10, wherein the second amino acid sequence further comprises a cleavable linker sequence between the purification tag and the amino acid sequence.

12. 10. The isolated peptide of claim 9, which is a fusion polypeptide comprising a first amino acid sequence of the peptide linked to a second amino acid sequence of the peptide.

13. A fusion polypeptide comprising a plurality of amino acid sequences linked together in series, each of said plurality of amino acid sequences comprising a peptide according to any one of claims 1 to 7.

14. A composition comprising one or more peptides according to any one of claims 1 to 6 or a fusion polypeptide according to claim 13 and a carrier.

15. The composition of claim 14 which is a clarified cell extract.

16. 15. The composition of claim 14, further comprising an additive selected from the group consisting of fertilizers, herbicides, insecticides, fungicides, nematicides, bactericides, biological inoculants, plant regulators, and mixtures thereof.

17. The additive is (i) clothianidin, a combination of clothianidin and Bacillus firmus, imidicloprid, or a combination of imidicloprid and Bacillus firmus, or (ii) Thiamethoxam; a combination of thiamethoxam, mefenoxam, and fludioxinil; a combination of thiamethoxam, mefenoxam, fludioxinil, and azoxystrobin; a combination of thiamethoxam and abamectin; a combination of thiamethoxam, abamectin, and a Pasteuria nematicide; or a combination of thiamethoxam, mefenoxam, fludioxinil, azoxystrobin, thiabendazole, and abamectin, or (iii) Biological inoculants including Bradyrhizobium sp., Bacillus sp., and combinations thereof. The composition of claim 16, comprising any one of

18. The composition of claim 14, wherein the carrier is an aqueous carrier.

19. 20. The composition of claim 18, wherein the aqueous carrier further comprises one or more of a biocide, a protease inhibitor, a non-ionic surfactant, or a combination thereof.

20. The composition of claim 14, wherein the carrier is a solid carrier in particulate form.

21. 21. The composition of claim 20, wherein the solid carrier is a dry powder.

22. applying an effective amount of the isolated peptide of any one of claims 1 to 7, the fusion polypeptide of claim 13, or the composition of claim 14 to the plant or plant seed or to the locus where said plant is growing or expected to grow. wherein said applying is effective to confer disease resistance.

23. applying an effective amount of the isolated peptide of any one of claims 1 to 7, the fusion polypeptide of claim 13, or the composition of claim 14 to the plant or plant seed or to the locus where said plant is growing or expected to grow. wherein said applying is effective to enhance plant growth.

24. applying an effective amount of the isolated peptide of any one of claims 1 to 7, the fusion polypeptide of claim 13, or the composition of claim 14 to the plant or plant seed or to the locus where said plant is growing or expected to grow. wherein said applying is effective to increase the resistance of said plant to a biotic stress factor selected from the group consisting of insects, arachnids, nematodes, weeds, and combinations thereof.

25. applying an effective amount of the isolated peptide of any one of claims 1 to 7, the fusion polypeptide of claim 13, or the composition of claim 14 to the plant or plant seed or to the locus where said plant is growing or expected to grow. wherein said applying is effective to increase the tolerance of said plant to an abiotic stress factor selected from the group consisting of salt stress, water stress, ozone stress, heavy metal stress, low temperature stress, high temperature stress, nutrient stress, and combinations thereof.

26. A DNA construct comprising a first nucleic acid molecule encoding the isolated peptide of any one of claims 1 to 7 or the fusion polypeptide of claim 13, and a promoter-effective nucleic acid molecule operably coupled to the first nucleic acid molecule.

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