BACKGROUND

[0001] Atrazine (2-chloro-4-ethylamino-6-isopropylamino-l,3,5-triazine) is an effective herbicide employed to control broadleaf and grass weeds, mainly in crops such as rice, wheat, maize, and sorghum. Because of its high stability, it is a well-known pollutant of great environmental concern. Atrazine has been shown to have negative effects such as DNA damage, gene expression shifts, cancer and endocrine disruption. Its residues are found in soil samples decades after it was last applied and were shown to chronically leach into local aquifers. As such, efforts are being made to limit and monitor its use.

[0002] While Atrazine was banned in the European Union and Switzerland since 2003, the United States Environmental Protection Agency still allows its wide monitored use. The areas contaminated with Atrazine and other hazardous herbicides (such as Diuron (3 -(3, 4- di chlorophenyl)- 1,1 -dimethylurea) and Linuron (3 -(3, 4-di chlorophenyl)- 1 -methoxy- 1- methylurea)) are rapidly increasing worldwide, introducing a need for remediation approaches. [0003] Bioremediation, an environmental bioprocess in which naturally occurring organisms are used for breaking down hazardous substances into less toxic or non-toxic substances, is increasingly acknowledged as a cost-effective feasible alternative for environmental cleaning. Critical environmental bioprocesses are naturally carried out by indigenous bacteria and are related to removal of pollutants from water, soil or air. Thus, native microbial bioremediation is widely applied for the biodegradation of various organic pollutants, including herbicides.

[0004] Bioremediation may occur naturally or, at a faster rate, by applying bio-stimulants, such as specific carbon or nitrogen sources, fertilizers, or electron acceptors, etc. These biostimulants, fertilizers, or electron acceptors encourage the growth of the degrading microbes within the medium or environment in a process termed bio-stimulation. Microbial metabolism is considered the most influential factor in Atrazine degradation, thus promoting the development of biodegradation strategies. One of the well-known Atrazine degrading bacteria are the gram-positive Paenarthrobacter aurescens Strain TCI (previously known as Arthrobacter aurescens Strain TCI), reported to biodegrade Atrazine into non-toxic components more efficiently than most other known degraders including Pseudomonas sp. strain ADP. The full hydrolytic pathway for Atrazine degradation in Paenarthrobacter aurescens strain TCI has been described.

[0005] Modern tools of genomics, transcriptomics, metabolomics, proteomics, signaling systems and synthetic biology have opened new avenues for biotechnological advances and are increasingly applied for promoting better understanding of complex biological systems. In recent years, constraint based metabolic modelling approaches have become widely used as an in-silico tool for organism-level phenotyping and the subsequent development of metabolic engineering strategies. Generally, such approaches follow four key steps: (1) data acquisition, mainly genome sequencing information, basic cell-physiological and biochemical knowledge and some experimental data on cell growth; (2) model reconstruction, translating data into structured mathematical representation; (3) constraint-based optimization simulations, the prediction of growth rate, substrate uptake rates, and byproduct rates under different growth conditions or following knockout mutations, in the absence of kinetic information; and (4) experimental validation. The potential of such models for the investigation of optimal processing is now acknowledged and practiced. Examples for applicative use include the optimal production or utilization of industrial compounds such as xylose, biofuels, vitamins and drug development.

[0006] In bioremediation, genome scale metabolic modeling approaches were applied for the design of the Geobacter sulfurreducens strain capable of increased electron transfer and higher Fe(III) respiration, beneficial for environmental bioremediation of uranium-contaminated groundwater. Application of genome scale metabolic modeling for strain design is often based on the “Optknock” algorithm and its derivatives which search for the optimal gene knockouts for a desired metabolic production. However, unlike industrial use of engineered strain for enhanced production in monoculture, introduction of exogenous species into natural habitat is far from trivial and hampers the application of modeling approaches towards bioremediation solutions (Scientific Reports, 2020, 10: 13019).

[0007] An alternative strategy is the use of modeling for the design of optimal conditions that enhance processes (degradation or synthesis) carried out by the endogenous community.

[0008] Commercial herbicides follow various mechanisms to disrupt the growth of the target plants. Most herbicides, including Atrazine, Diuron and Linuron, inhibit photosynthesis. Mainly they interfere with Photo System II (bind to the DI protein) in chloroplasts and hinder electron transport which stops the formation of high energy compounds, such as ATP. The reduced species further creates an unstable environment in the cell through the formation of reactive oxygen species, which leads to the oxidation of cellular lipids and affects the cell integrity. Thus, residual herbicides can impose undesirable crop growth constrains on future crops.

[0009] There is a need for beneficial compositions and methods to promote herbicide degradation. SUMMARY

[00010] The present invention provides, in one aspect, a method for (a) promoting the degradation of a herbicide in soil, (b) preventing a herbicide in soil from contaminating groundwater, or (c) preventing herbicide-related crop growth constrains, the method comprising the step of applying an effective amount of a bio-stimulant to herbicide-degrading bacteria in the soil, wherein the bacteria are not Arthrobacter sp. strain DAT1, or wherein or the bio-stimulant is not Glucose.

[00011] The present invention further provides, in another aspect, a method for stimulating herbicide-degrading bacteria, the method comprising the step of applying an effective amount of a bio-stimulant to the herbicide-degrading bacteria, wherein the bacteria are not Arthrobacter sp. strain DAT1, or wherein or the bio-stimulant is not Glucose.

[00012] In certain embodiments, the bacteria are Paenarthrobacter aurescens Strain TCI or Variovorax sp. strain SRS16.

[00013] In certain embodiments, the bacteria are Paenarthrobacter aurescens Strain TCI and the herbicide is Atrazine.

[00014] In certain embodiments, the bacteria are Variovorax sp. strain SRS16 and the herbicide is Linuron or Diuron.

[00015] In certain embodiments, the bio-stimulant is a disaccharide or an a-amino acid.

[00016] In certain embodiments, the bio-stimulant is a disaccharide, and the herbicide is

Atrazine.

[00017] In certain embodiments, the bio-stimulant is an a-amino acid and the herbicide is Linuron or Diuron.

[00018] In certain embodiments, the disaccharide is a di -Glucose.

[00019] In certain embodiments, the di -Glucose is Maltose or Trehalose.

[00020] In certain embodiments, the a-amino acid is Glutamine or Asparagine.

[00021] In certain embodiments, the herbicide comprises a Triazine, an aryl-urea, or a phenyl-urea.

[00022] In certain embodiments, the herbicide is selected from the group consisting of Atrazine, Diuron, and Linuron.

[00023] In certain embodiments, the effective amount of the bio-stimulant is at least about 5 ppm to at least about 2500 ppm.

[00024] In certain embodiments, the effective amount of Maltose is at least about 2500 ppm.

[00025] In certain embodiments, the effective amount of Trehalose is at least about 2500 ppm. [00026] In certain embodiments, the effective amount of Glutamine is at least about 20 ppm.

[00027] In certain embodiments, the effective amount of Asparagine is at least about 15 ppm.

[00028] In certain embodiments, the method further comprises sampling the soil for (a) the endogenous bacterial community, (b) the herbicide in the soil, or (c) the type of soil.

[00029] In certain embodiments, the method comprises promoting the degradation of Atrazine by applying an effective amount of Maltose to the soil.

[00030] In certain embodiments, the effective amount of Maltose is at least about 2500 ppm.

[00031] In certain embodiments, the method comprises promoting the degradation of Atrazine by applying an effective amount of Trehalose to the soil.

[00032] In certain embodiments, the effective amount of Trehalose is at least about 2500 ppm.

[00033] In certain embodiments, the method comprises promoting the degradation of Diuron by applying an effective amount of Glutamine to the soil.

[00034] In certain embodiments, the method comprises promoting the degradation of Linuron by applying an effective amount of Glutamine to the soil.

[00035] In certain embodiments, the effective amount of Glutamine is at least about 20 ppm.

[00036] In certain embodiments, the method comprises promoting the degradation of Linuron by applying an effective amount of Asparagine to the soil.

[00037] In certain embodiments, the effective amount of Asparagine is at least about 15 ppm.

[00038] In certain embodiments, the method comprises applying the bio-stimulant to the soil by (a) mixing a dry bio-stimulant with the soil, or (b) spraying a solubilized solution on the soil.

[00039] The present invention further provides, in another aspect, a bacteria biostimulant composition, comprising at least two bacteria bio-stimulants selected from the group consisting of a disaccharide or an a-amino acid.

[00040] In certain embodiments, the disaccharide is a di-Glucose.

[00041] In certain embodiments, the di-Glucose is Maltose or Trehalose.

[00042] In certain embodiments, the a-amino acid is Glutamine or Asparagine. [00043] In certain embodiments, the composition comprises (a) Maltose, (b) Trehalose, (c) Glucose and Maltose, (d) Glucose and Trehalose, (e) Maltose and Trehalose, or (f) Glucose, Maltose and Trehalose.

[00044] In certain embodiments, the composition is for stimulating (a) Paenarthrobacter aurescens Strain TCI, or (b) Variovorax sp. Strain SRS16.

[00045] In certain embodiments, the composition comprises (a) Maltose, (b) Trehalose, or (c) Maltose and Trehalose.

[00046] In certain embodiments, the composition is for stimulating Paenarthrobacter aurescens strain TCI.

[00047] In certain embodiments, the composition is for promoting the degradation of Atrazine in soil.

[00048] In certain embodiments, the composition comprises Glutamine.

[00049] In certain embodiments, the composition is for stimulating Variovorax sp. Strain SRS16.

[00050] In certain embodiments, the composition is for promoting the degradation of Diuron in soil.

[00051] In certain embodiments, the composition comprises (a) Glutamine, (b) Asparagine, or (c) Glutamine and Asparagine.

[00052] In certain embodiments, the composition is for stimulating Variovorax sp. Strain SRS16.

[00053] In certain embodiments, the composition is for promoting the degradation of Linuron in soil.

BRIEF DESCRIPTION OF THE FIGURES

[00054] Figure 1 is a bar graph illustrating the predicted and observed rate of atrazine degradation by different treatments (addition of different bio-stimulants to soil).

[00055] Figure 2 is a bar graph illustrating the predicted and observed rate of diuron degradation by different treatments (addition of different bio-stimulants to soil). None= no supplement; Glutamine = supplemented with glutamine as enhancer and Arginine = supplemented with arginine as inhibitor

[00056] Figure 3 is a line graph illustrating the rate of Bacterial biomass increase (A, B) and Linuron degradation (C, D) by different treatments (addition of different bio-stimulants to soil), as predicted (A, C) and experimentally observed (B, D). MS - linuron only. DETAILED DESCRIPTION

[00057] The present invention provides compositions and methods to initiate or expedite the degradation of herbicides. The inventors have surprisingly found, by first using elaborate computational prediction techniques, followed by reduction to practice in pot experiments, where applying different agents to the soil significantly enhanced the chemical breakdown of widely used commercial herbicides. Without being bound by any theory or mechanism, it is hypothesized that agents applied to soil act as bio-stimulants to endogenous bacterial strains and communities, thereby increasing their relative prevalence and/or activity. It is further hypothesized that this increase is responsible for the noted breakdown of the herbicides, either by promoting innate herbicide-degrading capabilities or by inducing such capabilities de-novo. Irrespective of any theory or mechanism, the present invention provides measurable, statistically-significant herbicide-degrading effects.

[00058] As exemplified herein, is has been surprisingly found that exemplary natural compounds, such as Maltose, Trehalose, Glutamine and Asparagine, significantly promote the degradation of exemplary commercial herbicides, such as Atrazine, Diuron and Linuron. These findings allow farmers and chemists, affordable and easy-to-use means for decreasing herbicide levels found, for example, in soil.

[00059] According to the principles of the present invention, these means may be used, i.e. applied to agricultural soil, at any stage of a crop’s growth cycle, in order to manipulate herbicide levels.

[00060] Further, according to the principles of the present invention, these means may be applied to soil once it is no longer used for growing crops, e.g. between growth cycles, in order to avoid potential undesirable effects of residual herbicides of previous growth cycles on seeds or crops of future growth cycles.

[00061] The present invention provides, in one aspect, a method for promoting the degradation of a herbicide, the method comprising the step of applying an effective amount of a bio-stimulant to herbicide-degrading bacteria.

[00062] The present invention provides, in another aspect, a method for preventing a herbicide from contaminating groundwater, the method comprising the step of applying an effective amount of a bio-stimulant to herbicide-degrading bacteria.

[00063] The present invention provides, in another aspect, a method for preventing herbicide-related crop growth constrains, the method comprising the step of applying an effective amount of a bio-stimulant to herbicide-degrading bacteria.

[00064] An artisan would appreciate that the expressions “promoting the degradation of a herbicide”, “preventing a herbicide from contaminating groundwater”, or “preventing herbicide-related crop growth constrains”, are relative to an untreated control, i.e. where a biostimulant was not applied to herbicide-degrading bacteria.

[00065] In certain embodiments, the herbicide is in soil. In certain embodiments, the herbicide-degrading bacteria are in soil. In certain embodiments, the herbicide is in soil and the herbicide-degrading bacteria are in soil.

[00066] An artisan would appreciate that the terms “degrade”, “degradation”, or “degrading” may encompass the process in which hazardous substances, including, in certain embodiments, herbicides, are broken down into less toxic or non-toxic substances. In certain embodiments, degradation of herbicides is bioremediation. In certain embodiments, degradation is biodegradation. In certain embodiments, degradation is achieved by naturally occurring organisms or microorganisms, which include, without limitation, bacteria.

[00067] An artisan would appreciate that the term “herbicide-degrading bacteria” may encompass one or more bacterial strains, bacterial cells, or bacterial cultures, capable of breaking down herbicides or weedkillers (such as chemical substances used to control unwanted vegetation). In certain embodiments, the bacteria are soil bacteria. In certain embodiments, the herbicide-degrading bacteria are soil bacteria. In certain embodiments, the soil is agricultural soil.

[00068] As used herein, the term "applying" refers to bringing in contact with a biostimulant of the present invention.

[00069] An artisan would appreciate that the term “bio-stimulant” may encompass substances and materials, which, when applied, encourage the growth of microorganisms, including, for example and without limitation, herbicide-degrading bacteria, or to increase their relative prevalence and/or activity. In certain embodiments, the bio-stimulant modifies microbial or bacterial metabolism in a way that increases herbicide degradation. In certain embodiments, the bio-stimulant modifies microbial or bacterial metabolism in a way that provides potential benefits to the growth and/or development of plants or crops.

[00070] In certain embodiments, the bio-stimulant of the present invention is applied in an effective amount. In certain embodiments, an “effective amount” is intended to include an amount of a bio-stimulant of the present invention alone or an amount of a combination of biostimulants claimed or an amount of a bio-stimulant of the present invention in combination with other active ingredients effective to act as stimulants. In certain embodiments, an "effective amount" of a bio-stimulant of the invention is that amount of bio-stimulant which is sufficient to provide a beneficial effect to the herbicide-degrading bacteria to which the bio-stimulant is applied. [00071] In certain embodiments, the bacteria are not Arthrobacter sp. strain DAT1. In certain embodiments, the bio-stimulant is not Glucose. In certain embodiments, the bacteria are not Arthrobacter sp. strain DAT1 and the bio-stimulant is not Glucose.

[00072] In certain embodiments, the herbicide is in soil, the herbicide-degrading bacteria are in soil, the bacteria are not Arthrobacter sp. strain DAT1, and the bio-stimulant is not Glucose.

[00073] In certain embodiments, the bacteria are Paenarthrobacter aurescens Strain TC 1 or Variovorax sp. strain SRS 16. In certain embodiments, the bacteria are Paenarthrobacter aurescens Strain TCI. In certain embodiments, the bacteria are Variovorax sp. strain SRS 16. In certain embodiments, the bacteria are Paenarthrobacter aurescens Strain TCI and Variovorax sp. strain SRS 16.

[00074] In certain embodiments, the bacteria are Paenarthrobacter aurescens Strain TCI and the herbicide is Atrazine.

[00075] In certain embodiments, the bacteria are Variovorax sp. strain SRS16 and the herbicide is Linuron or Diuron. In certain embodiments, the bacteria are Variovorax sp. strain SRS 16 and the herbicide is Linuron. In certain embodiments, the bacteria are Variovorax sp. strain SRS 16 and the herbicide is Diuron. In certain embodiments, the bacteria are Variovorax sp. strain SRS 16 and the herbicide is Linuron and Diuron.

[00076] In certain embodiments, the bio-stimulant is a disaccharide or an a-amino acid. In certain embodiments, the bio-stimulant is a disaccharide. In certain embodiments, the biostimulant is an a-amino acid. In certain embodiments, the bio-stimulant is a disaccharide and an a-amino acid.

[00077] In certain embodiments, the bio-stimulant is a disaccharide and the herbicide is Atrazine.

[00078] In certain embodiments, the bio-stimulant is an a-amino acid and the herbicide is Linuron or Diuron. In certain embodiments, the bio-stimulant is an a-amino acid and the herbicide is Linuron. In certain embodiments, the bio-stimulant is an a-amino acid and the herbicide is Diuron. In certain embodiments, the bio-stimulant is an a-amino acid and the herbicide is Linuron and Diuron.

[00079] In certain embodiments, the disaccharide is a di-Glucose.

[00080] In certain embodiments, the di-Glucose is Maltose or Trehalose. In certain embodiments, the di-Glucose is Maltose. In certain embodiments, the di-Glucose is Trehalose. In certain embodiments, the di-Glucose is Maltose and Trehalose. [00081] In certain embodiments, the a-amino acid is Glutamine or Asparagine. In certain embodiments, the a-amino acid is Glutamine. In certain embodiments, the a-amino acid is Asparagine. In certain embodiments, the a-amino acid is Glutamine and Asparagine.

[00082] In certain embodiments, the herbicide comprises a Triazine, an aryl-urea, or a phenyl-urea. In certain embodiments, the herbicide comprises a Triazine. In certain embodiments, the herbicide comprises an aryl-urea. In certain embodiments, the herbicide comprises a phenyl-urea. In certain embodiments, the herbicide comprises a Triazine, an aryl- urea, and a phenyl-urea.

[00083] In certain embodiments, the herbicide is selected from the group consisting of Atrazine, Diuron, and Linuron. In certain embodiments, the herbicide is Atrazine. In certain embodiments, the herbicide is Diuron. In certain embodiments, the herbicide is Linuron. In certain embodiments, the herbicide is Atrazine, Diuron, and Linuron.

[00084] In certain embodiments, the effective amount of the bio-stimulant is at least about 5 ppm. In certain embodiments, the effective amount of the bio-stimulant is at least about 50 ppm. In certain embodiments, the effective amount of the bio-stimulant is at least about 500 ppm. In certain embodiments, the effective amount of the bio-stimulant is at least about 2500 ppm. In certain embodiments, the effective amount of the bio-stimulant is at least about 5000 ppm.

[00085] In certain embodiments, the effective amount of the bio-stimulant is at least about 5 ppm to at least about 2500 ppm. In certain embodiments, the effective amount of the bio-stimulant is at least about 50 ppm to at least about 2500 ppm. In certain embodiments, the effective amount of the bio-stimulant is at least about 500 ppm to at least about 2500 ppm. In certain embodiments, the effective amount of the bio-stimulant is at least about 5 ppm to at least about 250 ppm. In certain embodiments, the effective amount of the bio-stimulant is at least about 5 ppm to at least about 25 ppm.

[00086] In certain embodiments, the effective amount of Maltose is at least about 25 ppm. In certain embodiments, the effective amount of Maltose is at least about 250 ppm. In certain embodiments, the effective amount of Maltose is at least about 2500 ppm.

[00087] In certain embodiments, the effective amount of Maltose is about 25 ppm. In certain embodiments, the effective amount of Maltose is about 250 ppm. In certain embodiments, the effective amount of Maltose is about 2500 ppm.

[00088] In certain embodiments, the effective amount of Maltose is about 250 ppm to about 25000 ppm. [00089] In certain embodiments, the effective amount of Trehalose is at least about 25 ppm. In certain embodiments, the effective amount of Trehalose is at least about 250 ppm. In certain embodiments, the effective amount of Trehalose is at least about 2500 ppm.

[00090] In certain embodiments, the effective amount of Trehalose is about 25 ppm. In certain embodiments, the effective amount of Trehalose is about 250 ppm. In certain embodiments, the effective amount of Trehalose is about 2500 ppm.

[00091] In certain embodiments, the effective amount of Trehalose is about 250 ppm to about 25000 ppm.

[00092] In certain embodiments, the effective amount of Glutamine is at least about 2 ppm. In certain embodiments, the effective amount of Glutamine is at least about 20 ppm.

[00093] In certain embodiments, the effective amount of Glutamine is about 2 ppm. In certain embodiments, the effective amount of Glutamine is about 20 ppm.

[00094] In certain embodiments, the effective amount of Glutamine is about 2 ppm to about 200 ppm.

[00095] In certain embodiments, the effective amount of Asparagine is at least about 1.5 ppm. In certain embodiments, the effective amount of Asparagine is at least about 15 ppm. [00096] In certain embodiments, the effective amount of Asparagine is about 1.5 ppm. In certain embodiments, the effective amount of Asparagine is about 15 ppm.

[00097] In certain embodiments, the effective amount of Asparagine is about 1.5 ppm to about 150 ppm.

[00098] In certain embodiments, the method further comprises sampling the soil for (a) the endogenous bacterial community, (b) the herbicide in the soil, or (c) the type of soil. In certain embodiments, the method further comprises sampling the soil for the endogenous bacterial community. In certain embodiments, the method further comprises sampling the soil for the herbicide in the soil. In certain embodiments, the method further comprises sampling the soil for the type of soil. In certain embodiments, the method further comprises sampling the soil for (a) the endogenous bacterial community, (b) the herbicide in the soil, and (c) the type of soil.

[00099] In certain embodiments, the method comprises promoting the degradation of Atrazine by applying an effective amount of Maltose to the soil.

[000100] In certain embodiments, the method comprises promoting the degradation of Atrazine by applying an effective amount of Trehalose to the soil.

[000101] In certain embodiments, the method comprises promoting the degradation of Atrazine by applying an effective amount of Maltose and Trehalose to the soil. [000102] In certain embodiments, the method comprises promoting the degradation of Diuron by applying an effective amount of Glutamine to the soil.

[000103] In certain embodiments, the method comprises promoting the degradation of Linuron by applying an effective amount of Glutamine to the soil.

[000104] In certain embodiments, the method comprises promoting the degradation of Diuron and Linuron by applying an effective amount of Glutamine to the soil.

[000105] In certain embodiments, the method comprises promoting the degradation of Linuron by applying an effective amount of Asparagine to the soil.

[000106] In certain embodiments, the method comprises promoting the degradation of Linuron by applying an effective amount of Glutamine and Asparagine to the soil.

[000107] In certain embodiments, the method comprises applying the bio-stimulant to the soil by (a) mixing a dry bio-stimulant with the soil, or (b) spraying a solubilized solution on the soil. In certain embodiments, the method comprises applying the bio-stimulant to the soil by mixing a dry bio-stimulant with the soil. In certain embodiments, the method comprises applying the bio-stimulant to the soil by spraying a solubilized solution on the soil. In certain embodiments, the method comprises applying the bio-stimulant to the soil by (a) mixing a dry bio-stimulant with the soil, and (b) spraying a solubilized solution on the soil.

[000108] In certain embodiments, the method comprises applying the bio-stimulant to the soil before applying plant or crop seeds to the soil. In certain embodiments, the method comprises applying the bio-stimulant to the soil after applying plant or crop seeds to the soil. In certain embodiments, the method comprises applying the bio-stimulant to the soil during growth of plant or crop in or on the soil. In certain embodiments, the method comprises applying the bio-stimulant to the soil before harvesting plant or crop from the soil. In certain embodiments, the method comprises applying the bio-stimulant to the soil after harvesting plant or crop from the soil.

[000109] The present invention further provides, in another aspect, a method for stimulating herbicide-degrading bacteria, the method comprising the step of applying an effective amount of a bio-stimulant to the herbicide-degrading bacteria.

[000110] An artisan would appreciate that the expression “stimulating herbicidedegrading bacteria”, is relative to an untreated control, i.e. where a bio-stimulant was not applied to herbicide-degrading bacteria.

[000111] In certain embodiments, the bacteria are not Arthrobacter sp. strain DAT1.

[000112] In certain embodiments, the bio-stimulant is not Glucose.

[000113] In certain embodiments, the bacteria are not Arthrobacter sp. strain DAT1 and the bio-stimulant is not Glucose. [000114] The present invention further provides, in another aspect, a bacteria biostimulant composition.

[000115] In certain embodiments, the composition comprises at least two bacteria biostimulants.

[000116] In certain embodiments, the bacteria bio-stimulants are selected from the group consisting of a disaccharide or an a-amino acid.

[000117] In certain embodiments, the disaccharide is a di-Glucose.

[000118] In certain embodiments, the di-Glucose is Maltose or Trehalose. In certain embodiments, the di-Glucose is Maltose. In certain embodiments, the di-Glucose is Trehalose. In certain embodiments, the di-Glucose is Maltose and Trehalose.

[000119] In certain embodiments, the a-amino acid is Glutamine or Asparagine. In certain embodiments, the a-amino acid is Glutamine. In certain embodiments, the a-amino acid is Asparagine. In certain embodiments, the a-amino acid is Glutamine and Asparagine.

[000120] In certain embodiments, the composition comprises (a) Maltose, (b) Trehalose, (c) Glucose and Maltose, (d) Glucose and Trehalose, (e) Maltose and Trehalose, or (f) Glucose, Maltose and Trehalose.

[000121] In certain embodiments, the composition comprises Maltose. In certain embodiments, the composition comprises Trehalose. In certain embodiments, the composition comprises Glucose and Maltose. In certain embodiments, the composition comprises Glucose and Trehalose. In certain embodiments, the composition comprises Maltose and Trehalose. In certain embodiments, the composition comprises Glucose, Maltose and Trehalose.

[000122] In certain embodiments, the composition is for stimulating (a) Paenarthrobacter aurescens Strain TCI, or (b) Variovorax sp. Strain SRS16. In certain embodiments, the composition is for stimulating Paenarthrobacter aurescens Strain TCI. In certain embodiments, the composition is for stimulating Variovorax sp. Strain SRS16. In certain embodiments, the composition is for stimulating (a) Paenarthrobacter aurescens Strain TCI, and (b) Variovorax sp. Strain SRS16.

[000123] In certain embodiments, the composition is for promoting the degradation of Atrazine. In certain embodiments, the composition is for promoting the degradation of Atrazine in soil.

[000124] In certain embodiments, the composition comprises Glutamine.

[000125] In certain embodiments, the composition is for promoting the degradation of Diuron. In certain embodiments, the composition is for promoting the degradation of Diuron in soil. [000126] In certain embodiments, the composition comprises (a) Glutamine, (b) Asparagine, or (c) Glutamine and Asparagine. In certain embodiments, the composition comprises Glutamine. In certain embodiments, the composition comprises Asparagine. In certain embodiments, the composition comprises Glutamine and Asparagine.

[000127] In certain embodiments, the composition is for promoting the degradation of Linuron. In certain embodiments, the composition is for promoting the degradation of Linuron in soil.

[000128] In certain embodiments, the composition comprises at least two bio-stimulants selected from the group consisting of (a) Maltose, (b) Trehalose, (c) Glucose, (d) Glutamine, and (e) Asparagine. In certain embodiments, the composition comprises at least three biostimulants selected from the group consisting of (a) Maltose, (b) Trehalose, (c) Glucose, (d) Glutamine, and (e) Asparagine. In certain embodiments, the composition comprises at least four bio-stimulants selected from the group consisting of (a) Maltose, (b) Trehalose, (c) Glucose, (d) Glutamine, and (e) Asparagine. In certain embodiments, the composition comprises (a) Maltose, (b) Trehalose, (c) Glucose, (d) Glutamine, and (e) Asparagine.

[000129] In certain embodiments, the composition comprises (a) Maltose, and/or (b) Trehalose, for promoting the degradation of Atrazine.

[000130] In certain embodiments, the composition comprises Glutamine, for promoting the degradation of (a) Diuron, and/or (b) Linuron.

[000131] In certain embodiments, the composition comprises (a) Glutamine, and/or (b) Asparagine, for promoting the degradation of Linuron.

[000132] When a range of values is expressed, embodiments include from the one particular value to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent "about, " it is understood that the particular value forms another embodiment. All ranges are inclusive and combinable. In some embodiments, the term “about”, refers to a deviance of between 0.0001-5% from the indicated number or range of numbers. In some embodiments, the term “about”, refers to a deviance of between 1-10% from the indicated number or range of numbers. In some embodiments, the term “about”, refers to a deviance of up to 25% from the indicated number or range of numbers.

[000133] The term “comprises” means encompasses all the elements listed, but may also include additional, unnamed elements, and it may be used interchangeably with the terms “encompasses”, “includes”, or “contains” having all the same qualities and meanings. The term "consisting of means being composed of the recited elements or steps, and it may be used interchangeably with the terms “composed of’ having all the same qualities and meanings. EXAMPLES

[000134] Decision support system (DSS) technology was applied in order to predict and validate bio-stimulants that are optimal for enhancing degradation of a herbicide/pollutant in soil.

[000135] Example 1. Maltose and Trehalose are bio-stimulants for Atrazine degradation.

[000136] Wheat (cv. “Jordan”) was used as the reporter plant based on its dose dependent sensitivity of shoot development performance (biomass and height) to atrazine concentration in soil. Experiments were carried out in replicates of five pots (0.5 L), 10 seeds sown in each. Soil from a non-herbicide treated field was mixed with bio-stimulants and sprayed with atrazine on soil surface. Degradation rate was estimated using a plant bioassay described in Xu, Zarecki et al. 2019 for atrazine degradation. Abundance of the suggested degrader species with and without bio-stimulant was detected using amplicon sequencing for the atrazine treated samples. Commercially available compounds were selected for validation. The selected compounds range in their predicted efficiency as bio-stimulants and include Trehalose and Maltose as strong enhancers, glucose as moderate enhancer, and OCDCA (Octadecanoic acid), serine and histidine as weak enhancers. The amount of carbon based bio-stimulants (maltose, trehalose, glucose, OCDCA) was applied according to Xu, Zarecki et al. 2019, when glucose at 15 g/Kg but not at 10 g/Kg induced a recovery of the reporter plant, based on the carbon content (6g /Kg of soil carbon supplemented). Nitrogen based bio-stimulants were used on the basis of normalized nitrogen, equivalent nitrogen (6.8 mg of nitrogen per pot) to supplemented atrazine in the control. In the calibration experiment, four concentrations of carbon based bio-stimulants and two concentrations of amino acids bio-stimulants were tested.

[000137] For each pot, soil was mixed with supplements using a cement machine (Shatal, 150 1), delivered into pots and sprayed with atrazine (500 g a.i. ha' ¹ ) on soil surface. Herbicides were applied using a motorized laboratory sprayer equipped with a flat fan nozzle (800 IE, Degania Sprayers Co. Ltd., Degania Bet, Israel) calibrated to deliver 300 1 ha' ¹ at 245 kPa, as described by Eizenberg et al., 2003. For each treatment, pots not treated with atrazine served as control, in addition to no-atrazine/no-supplement pots and atrazine/no-supplement pots. Pots were irrigated as needed by sprinklers. The experiment was carried out in a net house during July-August (Average temperature 28-32°C). The soils were collected after 15 days after recorded the effect of atrazine from all the pots and frozen (-80 °C) for further bacterial community analysis. [000138] All experiments were arranged in a completely randomized design. Effect of herbicide phytotoxicity was computed by one-way ANOVA. Means were compared by Tukey- Kramer honestly significant difference test (a, 0.05) using JMP software (vers. 7, SAS).

[000139] The simulations for the selected substrates are shown in Figure 1, for the recovery of a bioassay plant indicative of atrazine degradation in soil. Overall, the laboratory experiments supported predictions in the majority of the cases. Amplicon sequencing of the samples indicated a significant increase in the relative abundance of native species highly similar to Paenarthrobacter aurescens strain TCI in samples treated with the bio-stimulants Maltose and Trehalose, but not with other bio-stimulants.

[000140] Example 2. Glutamine is a bio-stimulant for Diuron degradation.

[000141] To study the impact of bio-stimulants on diuron degradation greenhouse experiments were carried out. The experimental design contained three groups based on supplements (None= no supplement; Glutamine = supplemented with glutamine as enhancer and Arginine = supplemented with arginine as inhibitor). Each group received two treatments, with diuron (containers filled with 10 Kgs of soil supplemented with 30ppm of diuron) and without diuron (containers filled with 10 Kgs of soil). The experiments for diuron were carried out in triplicate. The experiment was carried out in the green house at a temperature of ~25°C. [000142] The amount of diuron was detected with HPLC by following standard protocol. The supernatant was filtered through disposable 0.22 pm PTFE syringe filter and transferred to the HPLC vials for detection of residual diuron. Diuron was analyzed by using Agilent 1100 HPLC (Waldbronn Germany). Detection of diuron was done through UV absorption using the external calibration method (sensitivity 0.5 mg 1-1). The mobile phase with reverse phased Nova C18 column (Phenomenex Torrance, CA) of 250 mm length with particle size 5 pm was used for better separation. Diuron treated samples were sequenced for determining abundance of the suggested degrader species.

[000143] The amount of diuron was statistically analyzed for the differences in groups by using one way ANOVA at p<0.05 in rstatix package in R. The simulations for the Arginine and Glutamine are shown in Figure 2.

[000144] Example 3. Glutamine and Asparagine are bio-stimulants for Linuron degradation.

[000145] In vitro assays for determining the rate of degradation of Linuron by Variovorax sp. strain SRS16 were carried out using protocols reported in Ofaim, Zarecki et al. 2020. Commercially available compounds representing a range of compounds having enhanced predicted degradation potential and biochemical characteristics were selected for validation. These compounds include glutamine and asparagine as strong enhancers, aniline as a moderate enhancer, and arginine, oxalic acid, and catechol as weak enhancers and methionine whose enhancement potential depends on the choice of simulation pathway.

[000146] Variovorax sp. strain SRS 16 (NCBI: txid282217) was revived from stock cultures stored in glycerol at -80 °C. The purity and authenticity of the strain were checked by 16S rRNA gene sequencing. Bacterial cells were grown on minimal medium agar plates at 25 °C. For the quantitative analysis, 250 ml flasks were used to prepare minimal medium (50 ml of medium per flask) and autoclaved. The seven supplements (filter sterilized) were added to a final concentration of 0.12 mM in media (equivalent to 30 ppm of linuron). linuron (MS), MS+C & MS+N consist of only minimal medium, minimal media added with only carbon (glucose) and added with only nitrogen (ammonium salt), respectively. MS was treated as negative control. The autoclaved minimal medium was supplemented with the substrates (separately) and linuron (30 ppm) followed by bacterial inoculation to final OD (at 600 nm) of 0.05-0.1 of 2X MS washed mother culture. Mother culture was raised in MSCN (succinic acid and ammonium salt) medium with linuron, by inoculating fresh agar plate grown bacterial cells followed by incubation at 25 °C (120 rpm) for 24 h.

[000147] All the inoculated flasks were incubated at 25 °C (120 rpm) for seven days. Bacterial growth and linuron degradation were monitored at definite intervals (0 ^th Day, 3 ^rd Day, 5 ^th Day and 7 ^th Day) from the day of inoculation. For bacterial growth, 200 pl was taken and O.D. was measured at 600 nm by using Infinite® 200 PRO (Tecan Trading AG, Switzerland). Linuron degradation was measured through HPLC with standard procedures. Briefly, 1 ml of sample was taken and centrifuged 10000*g for 5 min. The supernatant was filtered through a 0.22 pm PTFE syringe filter and transferred to the HPLC vials for the detection of residual linuron. Linuron and 3, 4-dichloronaline were analyzed by using Agilent 1100 HPLC (Waldbronn Germany). Detection of linuron was done through UV absorption at 240 nm using the external calibration method (sensitivity 0.5 mgl' ¹ ). The mobile phase of 70% methanol at flow rate 1 min' ¹ with reverse phased Nova C18 column (Phenomenex Torrance, CA) of 250 mm length x 4.6 mm inner diameter with particle size 5 pm was used for good separation.

[000148] The effect of supplements was statistically analyzed by performing repeated ANOVA measures at p<0.05 in SPSS vl9.

[000149] The results (growth and degradation) for the seven selected substrates are shown in Figure 3 (Variovorax sp. strain SRS16). Overall, the laboratory experiments supported predictions in the majority of the cases. Growth experiments (Figure 3B) are fully consistent with predictions, showing a clear distinction between non-enhancer compounds and enhancers. In agreement with predictions, the experiments showed a slow (negligible) growth rate in linuron only (MS), and with MS supplemented by arginine, catechol, aniline and methionine. A significantly higher growth rate was recorded in the presence of glutamine and asparagine (p<0.05). Degradation results (Figure 3D) are overall consistent with growth results (Figure 3B) with significant stratification of enhancers (glutamine, asparagine) vs. non-enhancers (p<0.05). Aniline and methionine had a non-significantly stronger enhancement effect in comparison to MS, and were classified in the non-enhancers group (p<0.05 repeated measures ANOVA), consistent with simulations.

[000150] While certain features of the disclosure have been illustrated and described herein, many modifications, substitutions, changes, and equivalents may occur to those skilled in the art. It is, therefore, to be understood that the appended claims are intended to cover all such modifications and changes as fall within the true spirit of the disclosure.

REFERENCES

Ofaim, S., R. Zarecki, S. Porob, D. Gat, T. Lahav, Y. Kashi, R. Aly, H. Eizenberg, Z. Ronen and S. Freilich (2020). "Genome-scale reconstruction of Paenarthrobacter aurescens TCI metabolic model towards the study of atrazine bioremediation." Sci Rep 10(1): 13019.

Xu, X., R. Zarecki, S. Medina, S. Ofaim, X. Liu, C. Chen, S. Hu, D. Brom, D. Gat, S. Porob, H. Eizenberg, Z. Ronen, J. Jiang and S. Freilich (2019). "Modeling microbial communities from atrazine contaminated soils promotes the development of biostimulation solutions." ISME J 13(2): 494-508.

Eizenberg H, Goldwasser Y, Achdary G, Hershenhorn J. The potential of sulfosulfuron to control troublesome weeds in tomato. Weed Technol. 2003;17: 133-7.