EA045836B1 - PREVENTING OR REDUCING PLANT GROWTH BY BIOCEMENTATION - Google Patents

Description

The present invention primarily relates to the use of a mixture capable of biocementation as a means of preventing or reducing the growth of plants, in a preferred embodiment of the present invention, the growth of weeds, on/in a substrate. The invention also relates to a method for preventing or reducing the growth of plants, in a preferred embodiment of the present invention, the growth of weeds, on/in a substrate.

Other aspects and preferred forms of the present invention will be apparent from the following explanation, the accompanying examples and, in particular, the accompanying claims.

Uncontrolled growth of weeds is a constant problem in agriculture, in cities and municipalities, as well as in home gardens, as it leads to crop losses in agriculture, and gives a very unkempt and unattractive appearance to paths and other surfaces. For this reason, weeds are controlled and/or destroyed by thermal methods such as burning, manual weeding by hand or using various tools or chemicals. However, weeds are now becoming increasingly resistant to many existing products and therefore these products have to be reused, further increasing the cost of their use. In addition, treated areas are quickly repopulated by weeds due to the uncontrolled influx of seeds.

Therefore, the main object of the present invention was to provide a means for preventing or reducing unwanted plant growth, which would overcome the above-mentioned problems.

According to the present invention, this basic object is achieved by using a biocementable mixture as a means for preventing or reducing (through biocementation processes, respectively) plant growth, in a preferred embodiment of the present invention, weed growth.

As used herein, the term biocementation means essentially the natural thickening and/or hardening of (permeable) substrates (as defined below in the context of the method of the present invention). This prevents or reduces plant growth on/in these substrates. Biocement, within the scope of this text, is a product of biocementation as defined herein.

According to a preferred embodiment of the present invention, said essentially natural thickening and/or hardening is a process in which portions of the substrate on/in which plant growth is to be reduced or prevented are bound together by at least one binder contained in the mixture , to be used according to the present invention, and/or formed from its components, thereby thickening and/or hardening (biocementation) of the substrate or parts of the substrate. According to a particularly preferred embodiment of the present invention, said natural thickening and/or hardening is a process in which (living) organisms, parts thereof or enzymes, which, in a preferred embodiment of the present invention, are obtained from said organisms or parts thereof, and/ or produced by them are used to form carbonate, induce carbonate formation and/or catalyze carbonate formation. The carbonates formed bind parts of the substrate or substrate on/in which plant growth is to be reduced or prevented, resulting in thickening and/or hardening of the substrate or parts thereof. Therefore, the carbonates formed within the scope of this text, in the preferred embodiment of the present invention, represent a minimal component of the biocement. Optionally or in an alternative embodiment of the present invention, the addition of certain additives (as defined below) can (further) thicken and/or harden the substrate or portions of the substrate on which plant growth is to be reduced or prevented. According to another embodiment of the present invention, the biocement may also therefore consist of or contain additives or substances derived from them that are to be used according to the present invention (as defined below).

Therefore, according to a preferred embodiment of the present invention, removal of the substrate on/in which plant growth, preferably weed growth, is to be reduced or prevented is not necessary to prevent or reduce plant growth and therefore, in a preferred embodiment, is not part of the application. the present invention, especially since it is the thickening and/or hardening of the substrate (through biocementation processes, respectively) that prevents or reduces plant growth, in a preferred embodiment of the present invention, weed growth.

According to another preferred embodiment of the present invention, after removing the substrate on/in which plant growth, in a preferred embodiment of the present invention, weed growth, is to be reduced or prevented from its original location, said substrate is mixed with a mixture capable of biocementation (as described herein), in another location (for example, in a mixer) and (re)applying the resulting mixture at the original location (or, in an alternative embodiment of the present

- 1 045836 invention, in another location) where biocementation should be carried out.

Moreover, in the context of the application of the invention described herein, in a preferred embodiment of the present invention there is no need to compact the substrate on/in which plant growth is to be reduced or prevented, or the mixture of the substrate and the mixture capable of biocementation (as described in herein), or the formed biocement layer to achieve the prevention or reduction of plant growth, in a preferred embodiment of the present invention, weed growth, and thus, in a preferred embodiment of the present invention, this is also not a component of the application of the present invention.

In the context of this text, the term plant means terrestrial plants, that is, a monophyletic group of embryophytes characterized by a common, functionally understood complex of characters of several synapomorphies. Their main groups are: liverworts (Marchantiaceae), hornworts (Anthocerotaceae) and mosses (Bryopsidae), which are often grouped into the paraphyletic group of mosses, lycopodia (Lycopidae), horsetails (Equisetopsidae) and ferns in the narrow sense (Philicopsidae), as well as monophyletic seed plants plants (Speramotophytes) with angiosperms and various lineages derived from gymnosperms.

In the context of this text, the term weed means all plants (including mosses and ferns) of spontaneous or unwanted associated vegetation appearing among cultivated plants, in pastures or on (home) plots, which develops from the seed potential of the soil (as primary shoots or secondary shoots) , through root shoots, plant parts or seed tributaries, and which, in a preferred embodiment of the present invention, are not specifically cultivated there. Synonyms for the term weed are the terms wild plant and wild plant. In the context of this text, the term crop plants means plants whose growth is desirable.

The process of microbial biocementation is, for example, described in patent application WO 2006/066326 A1. The patent application discloses a process for forming a high-strength biocement in a permeable starting material wherein the starting material is added to a mixture with an effective amount of (i) a urease-producing microorganism, (ii) urea, and (iii) calcium ions. The urease-catalyzed conversion of urea to carbonate and its reaction with the supplied calcium ions produces calcium carbonate, which thickens the starting material. In particular, the process disclosed herein is described as being suitable for use in mining, civil engineering or specialty materials manufacturing.

Patent application WO 2016/010434 A1 describes a method for producing a self-healing cementitious material comprising mixing a cementitious starting material, a reducing agent and a fibrous reinforcing material, wherein the reducing agent comprises a bacterial material and the fibrous reinforcing material contains a biodegradable polymer. The bacteria used herein are capable of providing carbonates or phosphates, and they may be denitrifying bacteria, in accordance with one embodiment of the present invention.

The approach to solving the problem of the present invention is based on the use of biocementation to mechanically suppress, that is, to prevent or reduce the growth of plants, in the preferred embodiment of the present invention, the growth of weeds, by biologically thickening the substrate on/in which the plant grows. Therefore, the mixture to be used in the present invention is referred to below as a weed suppressant. Depending on the individual components of the mixture to be used according to the present invention (see below), in addition to mechanical inhibition, that is, preventing or reducing plant growth, other processes (as described below) may also play a role in inhibiting plant growth.

According to one embodiment of the present invention, the application of the mixture of the present invention described herein can be carried out in a locally limited area, thus in an area not treated with the mixture (as described herein) where thickening and/or curing is not carried out substrate on/in which plants grow, crop plants can (continue) to be cultivated.

According to another embodiment of the present invention, it is permissible to treat the substrate to be treated, on/in which plants grow, with a certain amount or type of mixture (as described herein) such that the resulting thickening and/or hardening of said substrate allows the growth of crop plants, but prevented or reduced weed growth. In this case, but also in general in connection with the present invention, the mixture to be used according to the present invention may also contain fertilizers that stimulate the growth of said crop plants.

Therefore, a preferred embodiment of the present invention relates to the use of the mixture of the present invention described herein, when the resulting thickening and/or hardening of the substrate on/in which the plant is growing allows the growth of at least one

- 2 045836 to the cultivated plant, but prevents or reduces the growth of weeds.

In an illustrative experiment, a field of arable soil was plowed and fertilized, kohlrabi seedlings were added to the arable soil, and the arable soil was intensively watered. The next day, the biocementable mixture (as described herein, see, for example, the examples section below) was applied to the arable soil either as a thickened mixture by hand or as a liquid mixture using a crop protection sprayer. In contrast to the control where water alone was used instead of a biocementable mixture (as described herein), good weed control was achieved with isolated weed germination while the crop plant (kohlrabi) continued to grow.

An advantage of the present invention is that the use of the mixture of the present invention for the purposes described herein results in a biocement layer thickness that is particularly suitable. In a preferred embodiment of the present invention, a biocement layer is obtained with a thickness of at least 1 mm, in a preferred embodiment of the present invention at least 3 mm, in a more preferred embodiment of the present invention at least 10 mm. In another preferred embodiment of the present invention, the layer thickness is required to be a maximum of 100 mm, in a preferred embodiment of the present invention a maximum of 50 mm, in another preferred embodiment of the present invention a maximum of 35 mm, in another preferred embodiment of the present invention a maximum of 30 mm.

As a result, in the most preferred embodiment of the present invention, the layer thickness of the entire formed biocement layer should be in the range from 1 mm to 100 mm, in the preferred embodiment of the present invention from 10 mm to 50 mm, in the more preferred embodiment of the present invention from 10 mm to 35 mm, in a more preferred embodiment of the present invention from 10 to 30 mm. The thickness of the biocement layer includes a section of the substrate, which thickens when the mixture is added. The thickness of the biocement layer can be determined by manual measurement after mechanical destruction of the layer using a gauge clamp. In an alternative embodiment of the present invention, various (non-destructive) measurement methods used in construction, agriculture, geology or other applications (eg, MIT-SCAN-T2 handheld device) can be used.

According to a preferred embodiment of the present invention, the use of the present invention mixture described herein results in the formation of a biocement layer that is permeable to water, that is, permeable or semi-permeable. This is a particular advantage of the present invention because, for example, rainwater can easily penetrate into the biocement layer and even flow into the biocement area. The water permeability of a sample is conventionally defined as the flow of water passing through the sample over a certain period of time. It may be expressed as a permeability rate (in cm/h, mm/h or cm/day) or alternatively as a permeability coefficient (in m/s). Specifying the permeability coefficient allows the sample, in a preferred embodiment of the present invention, a soil sample to be classified, for example, into the categories: (water)permeable, semi-permeable and (water)impermeable.

In the context of this text, the term permeable biocement layer means a biocement layer with a permeability coefficient (water) in the range from more than 10 ^-5 to 100 m/s, and the term water semi-permeable biocement layer means a biocement layer with a permeability coefficient (water) in the range from more than 10 ^{- 9} to 10 ^-5 m/s, and the term waterproof biocement layer is a biocement layer with a permeability coefficient (water) ranging from 10 ^-11 (or less) to 10 ^-9 m/s. Common methods for determining the permeability coefficient include laboratory methods (for example, probing a compacted core and then determining the water-saturated permeability in the laboratory) and field methods (for example, determining the infiltration rate using a two-ring infiltrometer).

A preferred embodiment of the present invention relates to the use of a mixture as defined herein, wherein the formed biocement layer has a permeability coefficient (water) ranging from more than 10 ^-9 to 100 m/s, in a preferred embodiment of the present invention from more than 10 ^{- 9} to 10 ^-3 m/s, in a more preferred embodiment of the present invention - from more than 10 ^-8 to 10 ^-3 m/s.

The use of the mixture of the present invention demonstrates reliable functionality under real-world (environmental) conditions, the mixture is easy to apply (often requiring only a single application), and it eliminates or reduces the use of chemical herbicide(s). In addition, it can be used in combination with existing weed control products or methods. The advantage of the present invention is that the application of the present invention is reversible, that is, the biocementation of the substrate or parts of the substrate can be reversed if necessary, for example, by the use of suitable acids or mechanical destruction, as well as by weathering and natural degradation. Thus, the substrate or parts

- 3 045836 substrate can again be available for the cultivation of cultivated plants.

Therefore, a preferred embodiment of the present invention relates to the use of a mixture as defined herein, wherein the biocementation of the substrate or parts of the substrate can be reversed or, in a preferred embodiment of the present invention, reversed.

Another preferred embodiment of the present invention relates to the use of a mixture as defined herein, wherein the mixture contains or consists of at least one organism and/or enzyme. In a preferred embodiment of the present invention, the use of the present invention relates to the use described above, wherein the mixture contains or consists of the following components:

(i) at least one organism and/or enzyme capable of forming carbonate and/or inducing and/or catalyzing carbonate formation, (ii) at least one substance for carbonate formation, (iii) optionally: at least one source cations; and (iv) optionally: at least one additive.

According to a preferred embodiment of the present invention, the organism or organisms in component (i) of the mixture to be used according to the present invention is/are one organism/organisms which, when performing a quantitative assay A, comprising the following steps:

Quantitative analysisA.

(i) providing and contacting the organism whose characteristics are to be obtained, or a mixture of organisms whose characteristics are to be obtained, with at least one carbonate-forming substance (and optionally other substances) and optionally a substrate, (ii) providing a means to determine ureolysis and/or carbonate formation, (iii) combining the mixture obtained in step (i) with the agent from step (ii), and (iv) determining the presence of ureolysis and/or carbonate formation using the agent from step (ii), leads to the determination of ureolysis and/or carbonate formation in step (iv), in the preferred embodiment of the present invention - if a substrate has been provided - to the determination of biocementation, in the preferred embodiment of the present invention - one that is sufficient to prevent or reduce plant growth, in a preferred embodiment of the present invention, weed growth (for more information on controlling the prevention or reduction of plant growth, see the examples section below).

An advantage of the present invention is that, in a quantitative assay A, both ureolytic and non-ureolytic organisms can be tested for their ability to adequately biocement.

The following explanations may be helpful in selecting organisms that are suitable for use in the present invention.

The carbonate-forming substance(s) that must be provided in step (i) of Quantitation A as defined herein, and the substrate that is thickened or solidified by biocementation that does not necessarily have to be provided, are further defined below, also with using selected and preferred examples. Optionally, other substances may be culture media, nutrient sources, cation sources (as described further below), and/or additives (as described further below).

In step (i) of quantitative analysis A, as defined herein, may be provided, for example, pure cultures of the organism to be characterized (for example, from strain collections), and/or the organism to be characterized, or a mixture of organisms , the characteristics of which are to be obtained can, for example, be isolated from a suitable sample (for example, from a soil sample) using a nutrient medium (for example, Christensen urea agar, B4 medium or M-3P medium) and cultured into a cell culture suitable for further research. The nutrient medium used for isolation and cultivation can be liquid or solid. One skilled in the art will know that, for example, the nutritional medium can be modified to suit the needs of the organism(s). The organism(s), in a preferred embodiment of the present invention, are cultured to a cell density in the range of 1 x 10 ⁷ to x 10 ¹² cells/ml. One skilled in the art will know that, for example, the culture temperature and the composition of the medium are selected according to the needs of the organism or mixture of organisms. The provided or prepared cell culture is then contacted with the substance(s) to form carbonate (and optionally other substances) and optionally with a substrate to produce a mixture, which is then combined in step (iii) with the agent from step (ii).

The means for determining ureolysis and/or carbonate formation in step (ii) of Quantitative Assay A, as defined herein, is, for example, a pH indicator, a device and/or at least one substance for measuring urease activity, a device and/or at least

- 4 045836 one substance for measuring the amount of carbonate formed by biocementation, or a device for measuring the degree of thickening of the substrate (by biocementation).

Determination of the presence of ureolysis and/or carbonate formation is present in step (iv) of quantitative analysis A as defined herein, in particular the determination of biocementation, in a preferred embodiment of the present invention - biocementation sufficient to prevent or reduce plant growth, in a preferred embodiment of the present invention, weed growth, may occur qualitatively or, in a preferred embodiment of the present invention, quantitatively.

A preferred method for such determination is, for example, adding a suitable pH indicator (eg phenol red, in a preferred embodiment of the present invention at a concentration of 15 mg/l) to the mixture leading to step (i). In the presence of ureolysis and/or carbonate formation, the pH of the mixture increases, resulting in a change in the color of the indicator (eg, a pink color in the case of phenol red).

If a source of cations, in the preferred embodiment of the present invention a source of calcium, is added to the mixture of step (i) (in addition to at least one carbonate-forming substance), then formation of a calcareous crust typically occurs around the colonies or on the colonies of the organism(s) in case of presence of ureolysis and/or carbonate formation in solid media. In the case of liquid culture media, limescale formation typically occurs if the cation source, in a preferred embodiment of the present invention a calcium source (eg CaCl2), and a carbonate source (eg urea) are available in sufficient quantities. Such formation of lime crust or lime deposit may also serve as optical evidence of the presence of ureolysis and/or carbonate formation, or said formation of lime crust or deposit may be analyzed by qualitative and/or quantitative determination of carbonate, in a preferred embodiment of the present invention with using Scheibler's (semi)quantitative carbonate determination or a similar but improved method (e.g., Horvath, B. et al., A Simple Method for Measuring Carbonate Content in Soils, Soil Science Society of America Journal 2005, 69, 1066 -1068).

Another preferred method for such determination is, for example, by measuring the urease activity of an organism or a mixture of organisms. Thus, the organism to be analyzed, or a mixture of organisms to be analyzed, is mixed, among other things, with a buffer urea (for example, 1.5 mol urea in 0.1 mol Tris-HCl, pH 7.5), and the formation of the formed ions ammonium is measured conductometrically as the measurement signal increases over time and urease activity is calculated (as described, for example, in BC Wiffin, Microbial Precipitation of CaCO ₃ for Biocement Production, Thesis, 2004, Murdoch University, Western Australia). Urease activity, in a preferred embodiment of the present invention, is in the range from 1x10'7 to 1x10'11 mmol hydrolyzed urea/min/cm/cells/ml, in another preferred embodiment of the present invention, in the range from 1x10'8 to 1x10' 10 mmol hydrolyzed urea/min/cm/cells/ml, in another preferred embodiment of the present invention in the range of 1x10'8 to 1x10'9 mmol hydrolyzed urea/min/cm/cells/ml. The first of the above corresponds to approximately a urea hydrolysis rate of 0-300 mmol hydrolyzed urea/min, depending on the number of cells used. Another preferred method for this determination is, for example, measuring the amount of carbonate formed by biocementation, in a preferred embodiment of the present invention using the Scheibler (semi-)quantitative determination of carbonate. The mixture to be tested, in a preferred embodiment of the present invention, is incubated for 48 hours in air at room temperature (25°C). The precipitated granule can then be obtained by centrifugation and drying for further use. The dried granule can be used for the (semi)quantitative determination of calcium carbonate formed, in a preferred embodiment of the present invention by Scheibler carbonate determination. Optionally, the dried sediment can be weighed in advance and the sedimentation efficiency can be calculated. Optionally, additional qualitative determination of ureolysis and/or carbonate formation can be performed in parallel. Phenol red (15 mg/l) can be added to the mixture from step (i). The supernatant, to be discarded when the pellet is removed, is then usually colored pink in the presence of ureolysis and/or carbonate pharmaceutics.

Another preferred method for this determination is, for example, by measuring the degree of thickening of the substrate (via carbonate obtained during biocementation). A suitable substrate for this is, for example, quartz sand, in a preferred embodiment of the present invention with a grain size of 0 to 2 mm (as a reference substrate). The remaining components of the mixture from step (i), in a preferred embodiment of the present invention, are applied to or introduced into the substrate in an amount of the resulting mixture of 5 l/m (in the case of a liquid mixture). Subsequent incubation should be carried out in air and at room temperature or above room temperature for at least 2 days (in the preferred embodiment of the present invention, at least 10 days). The strength of the formed biocement layer is then determined by mechanical failure analysis using a digital (fracture) force measuring device in accordance with DIN EN 196-1:2005-05. Compared to a control sample (using the comparison mixture without the organism or organisms on the substrate), differences in breaking force of >3 N (or >0.01 MPa) should be determined, in the preferred embodiment of the present invention, >30 N (or >0 ,1 MPa).

Within the scope of quantitative analysis A, the thickness of the biocement layer can also be determined using a gauge gauge; if thickening is successful, it should, in the preferred embodiment of the present invention, average >3 mm in the area of interest.

According to a preferred embodiment of the present invention, the organism or organisms in component (i) of the mixture to be used according to the present invention is/are one organism/organisms that, in step (iv) of quantitative analysis A, as defined herein with respect to , at least two determination methods described above, in a preferred embodiment of the present invention - at least three, in the most preferred embodiment of the present invention - at least four, in the most preferred embodiment of the present invention - all determination methods, leads (lead to) the determination of ureolysis and/or carbonate formation, in a preferred embodiment of the present invention - biocementation, in a preferred embodiment of the present invention - biocementation, which is sufficient to prevent or reduce plant growth, in a preferred embodiment of the present invention - weed growth (more for controls to prevent or reduce plant growth, see examples section below).

A preferred embodiment of the present invention is the use described above, wherein the mixture is present in liquid, gel, paste or powder form.

The mixture to be used in the present invention may be present or applied in the form of a liquid, gel, paste or powder mixture, or in the form of two, three, four or more liquid and/or gel and/or paste and/or powder pre-mixtures , which are present separately from each other, and which are mixed together before or during use.

An advantage of the present invention is that, in particular, the mixture or pre-mixes in powder form have a particularly long shelf life, in a preferred embodiment of the present invention from 12 to 24 months.

The powder form of the mixture or pre-mix can be prepared by standard methods known to one skilled in the art, for example, spray drying, freeze drying, (low temperature) vacuum drying, fluidized bed drying and/or filtration by filter media.

As used herein, the term powder means that the content of the liquid components, in the preferred embodiment of the present invention water, in the mixture is 10 wt.% or less, in the preferred embodiment of the present invention is 5 wt.% or less, in the preferred embodiment implementation of the present invention - 2.5 wt.% or less, in a more preferred embodiment of the present invention - 1.0 wt.% or less, in the most preferred embodiment of the present invention - 0.1 wt.% or less based on the total the mass of the mixture or pre-mix to be used according to the present invention.

The content of liquid components, in the preferred embodiment of the present invention - water, in the mixture or pre-mixture can be determined by standard methods known to one skilled in the art. For example, gravimetric determination of liquid components can be accomplished by weighing a sample, heating it to a temperature above the boiling point of the liquid components for a period of time sufficient to dry it, and then reweighing it. From the difference in weight before and after drying, it is possible to determine the content in % by weight of liquid components, in the preferred embodiment of the present invention - water.

According to another embodiment of the present invention, the mixture to be used according to the present invention may also be present or applied in the form of a gel or paste mixture or in the form of two, three, four or more separate solid and/or liquid and/or gel and/ or paste-like pre-mixes that are mixed together before or during use.

A preferred embodiment of the present invention is to be used as described above, wherein one, or all of the above mentioned organisms are/are selected from the group consisting of microorganisms, in a preferred embodiment of the present invention

- 6 045836 bretina, selected/selected from the group consisting of microorganisms of the phylum Firmicutes, in a preferred embodiment of the present invention - the class Bacilli, in a preferred embodiment of the present invention - the order Bacillales, in a preferred embodiment of the present invention - the families Planococcaceae or Bacillaceae, in in a preferred embodiment of the present invention, the genera Sporosarcina, Lysinibacillus or Bacillus, in a preferred embodiment of the present invention, are selected from the species Sporosarcina pasteurii, Sporosarcina ureae, Lysinibacillus sphaericus, Lysinibacillus fusiformis, Bacillus megaterium, Lysinibacillus sp., Bacillus pseudofirmus, Bacillus halodurans or Bacillus cohnii ; and from microorganisms of the phylum Proteobacteria, in a preferred embodiment of the present invention - the classes Alphaproteobacteria, Gammaproteobacteria, Deltaproteobacteria or Epsilonproteobacteria, in a preferred embodiment of the present invention - the orders Enterobacteriales, Myxococcales, Campylobacterales, Pseudomonadales or Caulobacterales, in a preferred embodiment of the present invention - the families Enterobacteriaceae, Myxococcaceae, Helicobacteraceae, Pseudomonadaceae or Caulobacteraceae, in a preferred embodiment of the present invention - the genera Proteus, Myxococcus, Helicobacter, Pseudomonas or Brevundimonas, in a preferred embodiment of the present invention, selected from the species Proteus vulgaris, Proteus mirabilis, Myxococcus xanthus, Helicobacter pylori, Pseudomonas aeruginosa or Brevundimonas diminuta; and from microorganisms of the phylum Actinobacteria, in a preferred embodiment of the present invention - the class Actinobacteria, in a preferred embodiment of the present invention - the order Actinomycetales, in a preferred embodiment of the present invention - the families Brevibacteriaceae or Micrococcineae, in a preferred embodiment of the present invention - the genera Brevibacterium or Micrococcaceae, in a preferred embodiment of the present invention, selected from the species Brevibacterium lines or Arthrobacter crystallopoietes; and from microorganisms of the phylum Cyanobacteria, in a preferred embodiment of the present invention - the class Cyanobacteria, in a preferred embodiment of the present invention - the order Synechococcales, in a preferred embodiment of the present invention - the family Synechococcaceae, in a preferred embodiment of the present invention - the genus Synechococcus, in a preferred embodiment of the present invention - Synechococcus species; and from aerobic bacteria, anaerobic bacteria, facultative anaerobic bacteria and intermediate species thereof.

This includes all variants, serotypes, mutants and spores, as well as any derivatives of genetically modified microorganisms.

The above organism(s), in a preferred embodiment of the present invention microorganisms, may be present (together or separately) in liquid(s), such as buffers, solvents, culture media and/or mixtures thereof, which may also be frozen at low temperatures. temperature, or they may be present in powder form.

According to the present invention, an organism or organisms capable of forming carbonate and/or inducing and/or catalyzing carbonate formation is or is part of the mixture used.

In an alternative embodiment of the present invention, it is permissible and disclosed in the context of this text that the (indigenous) organisms present in the substrate, in the preferred embodiment of the present invention, in the soil in/on which plant growth occurs, or isolated from said substrate , cultivated in laboratory conditions and then reintroduced onto/into the substrate, are capable of forming carbonate and/or inducing and/or catalyzing carbonate formation. In this case, alternative or equivalent embodiments of the present invention are possible (and accordingly included herein in accordance with the present invention), in which the organisms of the mixture to be used according to the present invention and the (indigenous) organisms in/on the substrate together form carbonate, induce and/or catalyze carbonate formation, or in which the mixture to be used does not itself contain organisms capable of forming carbonate, inducing and/or catalyzing carbonate formation. According to a preferred embodiment of the present invention, component (i) of the mixture to be used according to the present invention contains or consists of a combination of one organism or more organisms capable of forming carbonate, inducing and/or catalyzing carbonate formation, and organisms that are not capable of doing so .

According to a preferred embodiment of the present invention, component (i) of the mixture to be used according to the present invention contains or consists of a combination of aerobic bacteria, anaerobic bacteria and/or facultative anaerobic bacteria, and/or intermediate species thereof.

According to another preferred embodiment of the present invention, component (i) of the mixture to be used according to the present invention contains or consists of a combination of one or more organisms capable of forming carbonate in a ureolytic manner, inducing and/or catalyzing carbonate formation in a ureolytic manner, and organisms not capable of ureolysis, or not capable of forming carbonate and/or inducing, and/or

- 7 045836 catalyze carbonate formation.

Thus, one skilled in the art will know that biocementation (as defined herein) is particularly effective over a certain range of cell counts of the organisms used when using the mixture (as defined herein) of the present invention. According to our own research, the number of cells of the organism or organisms in the mixture to be used according to the present invention is, in a preferred embodiment of the present invention, at least 10 ⁷ cells/ml, in a more preferred embodiment of the present invention, at least 10 ⁸ cells/ml and/or, in a preferred embodiment of the present invention, at least 10 ¹² cells/ml, in a more preferred embodiment of the present invention, at least 10 ¹⁰ cells/ml, in a most preferred embodiment of the present invention, at least at least 10 ⁹ cells/ml. According to a preferred embodiment of the present invention, the number of cells of the organism or organisms in the mixture to be used according to the present invention is from 10 ⁸ to 10 ⁹ cells/ml.

The use described above is preferred, wherein one or more or all of the above enzymes are/are selected from the group consisting of urease, asparaginase, carbonic anhydrase and metabolic enzymes.

Metabolic enzymes in the context of this text, in a preferred embodiment of the present invention, are metabolic enzymes of at least one (micro)organism described herein, which, for example, by converting acetate and/or lactate, is capable of forming carbonate and/or induce and/or catalyze carbonate formation. In a preferred embodiment of the present invention, at least one organism (as defined above) capable of producing at least one of the above enzymes is used in component (i) of the mixture to be used according to the present invention, or, in a preferred embodiment of the present invention invention, the above enzymes are obtained or released from the above organisms.

If the organism or organisms used are pathogenic organisms, then in a preferred embodiment of the present invention, within the scope of this text, only non-pathogenic enzymes that are obtained or released therefrom are used in component (i) of the mixture to be used according to the present invention.

According to another preferred embodiment of the present invention, combinations of enzymes obtained or released from the above-mentioned organisms with enzymes of non-microbial origin (eg, plant enzymes) can be used in component (i) of the mixture to be used according to the present invention. For example, the urease enzyme can be obtained from soybeans and used in the present invention.

According to another preferred embodiment of the present invention, a combination of at least one of the abovementioned organisms capable of forming carbonate and/or inducing and/or catalyzing carbonate formation with at least one of the abovementioned enzymes capable of forming carbonate and/or inducing, and/or catalyze carbonate formation, can be used in component (i) of the mixture to be used according to the present invention.

Carbonates can be produced through various metabolic processes using the above mentioned enzymes. For example, aerobic metabolism of organic carbon sources may result in ammoniation (eg, the enzyme asparaginase) or heterotrophic metabolism of organic carbon sources may occur (eg, calcium lactate or calcium acetate). Both of these processes result in the formation of carbonates. Aerobic and anaerobic photosynthesis can also be used to form carbonates, as can anaerobic denitrification, anaerobic sulfate reduction and (an)aerobic methane oxidation.

Thus, biocementation using the mixture to be used according to the present invention can be based on at least one of the above metabolic processes.

Therefore, the use described above is preferred, wherein one, more or all of the above-mentioned carbonate-forming substances are selected from the group consisting of urea and its salts, organic acids such as lactic acid and its salts, in a preferred embodiment implementation of the present invention - carboxylates and their esters, gluconic acid and its salts, in a preferred embodiment of the present invention - carboxylates and their esters, acetic acid and its salts, in a preferred embodiment of the present invention - carboxylates and their esters, formic acid and its salts , in a preferred embodiment of the present invention - carboxylates and their esters, peptides, in a preferred embodiment of the present invention - containing asparagine, glutamine and/or glutamic acid, amino acids, in a preferred embodiment of the present invention - asparagine, glutamine and glutamic acid and its salts , in a preferred embodiment of the present invention

- 8 045836 nia - carboxylates and their esters, plant and animal complex substrates, in particular peptone, yeast extract, meat extract, nutrient broth and casamino acid, industrial waste streams, in particular corn extract, lactose mother liquor, protein lysates, in a preferred embodiment of the present invention - from peas, meat or tomatoes, anaerobic substrates, in a preferred embodiment of the present invention - from carbon dioxide and methane.

In addition, a preferred embodiment of the present invention is the use described above, wherein one, more or all of the above cation sources are/are selected from the group consisting of organic and inorganic calcium salts, in a preferred embodiment of the present invention calcium nitrate, acetate calcium, calcium lactate and calcium chloride, magnesium salts, manganese salts, zinc salts, cobalt salts, nickel salts, copper salts, lead salts, iron salts, cadmium salts, polymers, in a preferred embodiment of the present invention - cationic polymers, heavy metal cations , light metal cations, radioactive cations and their mixtures.

According to the present invention, the cation source(s) may or may not be contained in the mixture to be used according to the present invention. If they are not contained in the mixture, they may be present in/on the substrate or added to the substrate on/in which the plant is growing to allow bio-cementation.

A preferred embodiment of the present invention is also to be used as described above, wherein one, more or all of the above additives are selected from the group consisting of the following substances/mixtures of substances (in a preferred embodiment of the present invention, until it /they will no longer be contained in component (ii)): nutrients; (bio)polymers, in a preferred embodiment of the present invention - polyhydroxybutyrate (PHB), polylactide (PLA), polybutylene succinate (PBS), polyacrylic acid (PAA), polymethacrylate (PMA), poly(2-hydroxyethyl methacrylate) (PHEMA), polyvinyl alcohol (PVOH), polyvinyl acetate (PVAC), polyvinylpyrrolidone (PVP), poly(2-ethyl-2-oxazoline), polystyrene (PS), polyamide, copolymers, polyamic acids, cellulose and its derivatives, starch and its derivatives, lignins and their derivatives, pectins and their derivatives, natural glue, in particular gum arabic, latex, rubber and their derivatives, chitin and its derivatives, chitosan and its derivatives, cyclodextrins and their derivatives, dextrins and their derivatives; hydrogel formers, in a preferred embodiment of the present invention xanthan gum, alginates and agar-agar; cold-soluble and/or heat-soluble (vegetable) glue; calcium carbonate and mixtures containing calcium carbonates, in a preferred embodiment of the present invention - nacre, amorphous calcium carbonate, precipitated calcium carbonate, aragonite, calcite, vaterite and mixtures and derivatives thereof; polysaccharides and extracellular polymeric substances (EPS), in a preferred embodiment of the present invention - microbial exopolysaccharides, in a preferred embodiment of the present invention, containing or consisting of maleic acid, acetic acid, lactic acid, lactose, sucrose, glucose, fructose and/or inulin ; sources of protein, fibers and fibrous materials, in a preferred embodiment of the present invention - casein, albumin, yeast extracts, peptones, cellulose fibers, wood fibers, wood-cellulose fibers; residues and industrial materials, in a preferred embodiment of the present invention, corn extract, lactose mother liquor, protein lysates, molasses, protein waste, in a preferred embodiment of the present invention, from yeast production, meat production, dairy industry and paper production; silicates and their derivatives; acrylates and their derivatives; wasserglass and binders like wasserglass; cement and cement additives, in the preferred embodiment of the present invention - sand, lime and their derivatives, aluminum oxide, calcium oxide, calcium hydroxide, aluminum hydroxide, ash, in the preferred embodiment of the present invention fly ash and bone ash, microsilica, kaolin, bentonites , fillers, in a preferred embodiment of the present invention - white lime (hydrate), crushed limestone sand and limestone powder; resins and epoxides; natural and chemical herbicides; fungicides; molluscicides; insecticides; water repellents and wax emulsions; emulsifiers; binders; thixotropic agents; crystallization nuclei and crystallization modifiers; fatty acids; minerals and trace elements; salts, in a preferred embodiment of the present invention - phosphates and sulfates; rocks, in a preferred embodiment of the present invention pumice and shale powder; bacteria capable of forming polymers; and biocementation modifying substance(s).

In the context of the present invention, the additive(s), in a preferred embodiment of the present invention, are either substances that affect the biocementation process itself (for example, nutrients), or substances that affect the final product of biocementation, that is, the properties of the biocement (for example , its water resistance), or substances affecting plant growth

- 9 045836 which is suppressed (for example, herbicides).

Examples of substances that can influence the properties of biocement, in a preferred embodiment of the present invention, are additives to be used according to the present invention, which are capable of (further) thickening and/or hardening of the substrate or parts of the substrate on which plant growth is to be reduced or prevention. These may be, for example, sugar molecules (as defined above) or polymers formed by bacteria. In this case, as described above, the additives to be used in the present invention are, if applicable, the sole component of the biocement (alternatively or in addition to the formed carbonate(s) of the preferred embodiment of the present invention).

According to the present invention, the additive(s) may be present in the mixture to be used according to the present invention (ie, be part of the mixture to be used) or contained in the substrate on which the plant grows. In an alternative embodiment of the present invention, they may also not be present.

In the context of the present text, monomers of the additives mentioned above, in particular the mentioned (bio-)polymers, are also considered as additives to be used in the present invention.

One skilled in the art will understand that the number of uses and mode of action of the additive(s) will largely depend on its own properties or on the properties of other components of the mixture to be used according to the present invention or on the properties of the substrate, and will select accordingly suitable combinations and number of applications of the additive(s).

A preferred embodiment of the present invention is also the use described above, wherein the plant or weed is selected from the group consisting of dicotyledonous plants of the following genera: Abutilon, Gooseberry, Kokorysh, Shchiritsa, Ambrosia, Anhusa, Anagallis, Anode, Antemis, Nevzrachnitsa, Arabidopsis , Quinoa, Crescent, Daisy, String, Sverbiga, Shepherd's purse, Thistle, Cassia, Cornflower, Goosefoot, Chrysanthemum, Thistle, Hemlock, Conysa, Consolidum, Bindweed, Datura, Descurainia, Desmodium, Emex, Horsetail, Erigeron, Erodium, Jaundice, Euphorbia, Furmaria, Pikulnik, Galinzoga, Bedstraw, Geranium, Hogweed, Hibiscus, Morning Glory, Kochia, Sweetweed, Warthog, Lathyrus, Lepidium, Lithospermum, Linaria, Lindernia, Crookedflower, Mallow, Matricaria, Mint, Woodweed, Mollugo, Forget-me-not, Papaver, Farbitis, Plantain, Polygonum, Purslane, Ranunculus, Radish, Zherushnik, Rotala, Sorrel, Solyanka, Rasson, Sesbania, Sida, Synapis, Gooseberry, Solanum, Sow thistle, Sphenoclea, Stachys, Chickweed, Dandelion, Jarutka, Clover, Mother-and- stepmother, Nettle, Veronica, Viola, Cockurberry; dicotyledonous plants of the following genera: Peanut, Beetroot, Brassica, Cucumis, Pumpkin, Helianthus, Carrot, Wisteria, Cotton, Morning Glory, Lettuce, Flax, Tomato, Tobacco, Beans, Peas, Solanum, Pea monocotyledonous plants of the following genera: Aegilops, Zhitnyak, Bentgrass , Alopecurus, Metlitsa, Oats, Brachiaria, Bonfire, Cenchrus, Commelina, Pigwort, Cyperus, Dactyloctenium, Crabgrass, Barnyardgrass, Eleocharis, Eleusina, Eragrostis, Woolgrass, Fescue, Fimbristylis, Heteranthera, Imperata, Murraya, Juncus, Leptochloa, Tares, Monochoria , Panicum, Paspalum, Canary grass, Timothy grass, Bluegrass, Rothbellia, Sagittaria, Scirpus, Setaria, Sorghum; and monocotyledonous plants of the following genera: Allium, Pineapple, Asparagus, Oats, Barley, Rice, Panicum, sugar cane, Rye, Sorghum, Triticale, Wheat, Corn; mosses of the class liverworts (Marshantiaceae), hornworts (Anthocerotaceae), mosses (Bryopsidae);

in a preferred embodiment of the present invention, the growth of at least two, three, four, five, six, seven, eight, nine, ten, more than ten or all of these plants is prevented or reduced.

According to a preferred embodiment of the present invention, one, more or all of the above plants are at least one liverwort selected from the group consisting of the following genera: Acolea, Acrobolbus, Acrochilla, Acromastigum, Acroscyfella, Acrocyphus, Acrostolia, Adelocolia, Aychisoniella, Alicularia, Allisonia, Allisoniella, Alobiella, Alobiellopsis, Amazoopsis, Amphicephalosia, Amphilophocolea, Andrewsianthus, Aneura, Anomacaulis, Anomoklada, Anomylia, Anthelia, Antelis, Aphanolezhenea, Aplosia, Apomarsupella, Apometsgeria, Apotreubia, Arachniopsis, Arctosciphus, Arnellia, Ascidiota, Asterella, Athalamia, Austrofossombronia, Austrolembidium, Austrolophosia, Austromezgeria, Austroscifus, Balantiopsis, Bazzania, Blasia, Blefaridophyllum, Blepharostoma, Brevianthus, Calicularia, Calypogea, Calyptrocolea, Campanocolea, Castanoclobos, Cavicularia, Cephaloonesia, Cepha lolobus, Cephalomitrion, Cephalosia, Cephalosiella, Cephalosiopsis, Keratolezhenea, Caesius, Chaetophyllopsis, Chiastocavlon, Hyloscyphus, Chloranthelia, Honecolea, Cladomastigum, Cladopodiella, Clandarium, Clasmatocolea, Cololezhenea, Kolura, Conocephalum, Conoscyphus, Corsinia, Kronizia, Crossogyna, Cryptochila, Cryptocolea, Cryptocolepsis, Crypt Omitrium, Cryptostipula, Cryptothallus, Cuspidatula, Cyanolophocolea, Cyathodium, Cylindrocolea, Delavayella, Dendrobazania, Dendromastigophora, Denotarysia, Dichiton, Dinkleria, Diplocolea, Diplophyllum, Duinia, Drepanolea, Drucella, Dumortiera, Dumortieropsis, Enigmella, Eocalypogeia, Eoisotachis, Eopleurosis, Eotrichocole I, Eremonotus, Eucalyx,

- 10 045836

Evansia, Evansianthus, Exormotheca, Fossombronia, Frullania, Fuscocephalosiopsis, Gaxtremia, Geocalix, Geothallus, Gerhildiella, Goebeliella, Goebelbrium, Gongilanthus, Gottshea, Gottschelia, Greeneothallus, Grollea, Gymnante, Gymnocoleopsis, Gymnomitrion, Gymnoscyphus, Gyrothyra , Hesselia, Haplomitrium, Harpalezhenea, Harpanthus, Gattoria, Gattorianthus, Gattoriella, Hepatostolonophora, Herbertus, Herpetium, Herpocladium, Herzogianthus, Herzogobrium, Heterogemma, Heteroscyphus, Horikavella, Hyalolepidosia, Hygrobiellalvatsukia, Hygrolembidium, Hygrophila, Hymenophyton, Hypoisotachis, Isolembidium, Iso takis, Jamsoniella, Jensenia, Yubula, Yubulopsis, Jungermannia, Jungermannites, Crunodiplophyllum, Curzia, Cymatocalyx, Lamellocolea, Leiocolea, Leiomithra, Leiomilia, Leioscyphus, Lezhenea, Lembidium, Lepidogyna, Lepidolena, Lepidozia, Leptoleugea, Leptophyllopsis, Leptoscyphopsis, Leptoscyphus, Letocolea, Liochlena, Lo bathyriccardia, lophocolea, lophonardia, lofosia, Lophosiopsis, Lunularia, Macrodiplophyllum, Maculia, Mackinoa, Mannia, Marchantia, Marchesinia, Marsupella, Marsupidium, Massula, Massularia, Mastigobrium, Mastigopelma, Mastigophora, Mastigopsis, Mesoptychia, Metacalypogeia, Metaigrobiella, Metzgeria, Metzgeriopsis, Micrizophylla, Microle Zhenya, Microlepidosis, Micropterygium, Misutania, Mnioloma, Murkia, Monocarpus, Monoclea, Monodactylopsis, Monosolenium, Mytilopsis, Nanomarsupella, Nardia, Neesioscyphus, Neogrollea, Neochodzonia, Neotrichocolea, Noteroclada, Notogymnomitrion, Notostrepta, Notscyphus, Novellia, Obtusifolium, Odon tolerea, Odontoschism, Oleolophosia, Oximitra, Pachyglossa, Pachyschistochila, Pallavicinia, Paracromastigum, Paraschistochila, Patarola, Pedinophyllopsis, Pedinophyllum, Pellia, Peltolepsis, Perdusenia, Perssoniella, Petalophyllum, Phycolepidosia, Phyllotallia, Physiothium, Physoteca, Pisanoa, Plagiochasma, Plagiochilla, Plagiochilidium, Plagiochilion , Platykaulis, Plectocolea, Pleurante, Pleuroclada, Pleurocladopsis, Pleurocladula, Pleurosia, Podante, Podomitrium, Porella, Prasanthus, Preissia, Prionolobus, Protolophosia, Protomarsupella, Protosysgiella, Protosyzygiella, Pseudocephalosia, Pseudocephalosiella, Pseudolophocolea, Pseudolophosia, Pseudomarsupidium, Pseu donevra, Pseudotritomaria, Psiloklada, Pteropsiella, Ptilidium, Radula, Rebulia, Rhizocaulia, Rhodoplagiochila, Riccardia, Riccia, Ricciella, Ricciocarpos, Riella, Roivainenia, Ruisanthus, Ruttnerella, Saccobasis, Saccogina, Sandeothallus, Sarcocyphos, Sarcomitrium, Sauteria, Scapania, Scaphophyllum, Schiffneria, Schisma, Schistochila, Schistochilaster, Schistochillo dog, Scofildia, Sendtner, Seppeltia, Sevardiella, Simodon, Solenostoma, Sothbia, Spherocarpus, Sphagnoecetis, Sprucella, Steereella, Stereocolea, Stenorripis, Stephandium, Stephiniella, Stefaniellidium, Stephensoniella, Symphyogyna, Symphiogynopsis, Symphiomithra, Synchimenium, Sizgiella, Teniolezhenea, Targio niya, Tegulifolium, Telaranea, Tallocarpus, Treubia, Triandrophyllum, Trichocolea, Trichocoleopsis, Trichostilium, Trichothemnoma, Trilophosia, Tritomaria, Tilimanthus, Vanaea, Vandiemenia, Verdornia, Vetaforma, Vettsteinia, Wiesnerella, Xenochila, Xenothallus, Zoopsidella, Zoopsis.

According to another preferred embodiment of the present invention, one, more or all of the above plants are at least one moss selected from the group consisting of the following genera: Abietinella, Acanthocladiella, Acanthocladium, Acanthodium, Acanthorhynchium, Akavlon, Akavlonopsis, Achrophyllum, Acidodontium , Acrocladium, Acroporium, Acroschisma, Actinodontium, Actinotuidium, Adelothecium, Equatorella, Erobryidium, Erobryopsis, Erobrium, Erolindigia, Algaria, Aligrimmia, Alleniella, Allioniellopsis, Aloina, Aloinella, Alofosia, Alzia, Amblyodon, Amblyodum, Amblystegiella , Amblystegium, Amblytropis, Ambukhanania , Amphidium, Amphoridium, Amphoritheca, Anakalipta, Anakamptodon, Anacolia, Ancistrodes, Andoa, Andreea, Andreobrium, Aniktangium, Anisothecium, Anodon, Anodontium, Anoectangium, Anomobrium, Anomodon, Antitrichia, Ongstremia, Ongstremiopsis, Apalodium, Aphanorregma, Apio carp, Aplodon, Apterygium . lacomnium, Aulacopylum, Avstinella, Austrochondella . cium, Brachytrichum, Braithwaitea , Brunfelsia, Braunia, Breidleria, Breutelia, Brotera, Broterella, Broterobrium, Bruchia, Brychnia, Brimela, Bryoandersonia, Bryobeckettia, Bryobrittonia, Bryobrotera, Bryoceutospora, Bryochenea, Bryokrumia, Bryodixonia, Bryodosenia, Bryoerythrophyllum, Bryohaplocladium, Bryohumbertia, Brie Omalthea, Bryomaginia, Bryomnium , Bryonoguchia, Bryonorrhizia, Bryopyxia, Bryozedgvikia, Bryostreymannia, Bryotestua, Bryum, Bukiella, Buklandiella, Burnettiia, Buxbaumia, Callialaria, Callicladium, Callicosta, Callicostella, Callicostellopsis, Calliergidium, Calliergon, Caloghypnum, Kalimperastrum, Kalimperes, Kalimper Idium, Kalimperopsis, Calyptopogon, Calyptothecium , Calyptrochaete, Camptochaete, Camptodontium, Camptothecium, Campiliadphus, Campyldium, Campilium, Campylodontium, Campylophyllum, Campylopodiella, Campylopodium, Campylopus, Campylostelium, Canalopopterygium, Cardocia, Cardocilla, Caribeohypnum, Catagoniopsis, Catagonium, Catarinea, Catarinella, Catharomnion, Katoscopium, Cecalifum, Ceratodom , Ceutospora, Ceutoteca, Heto

- 11 045836 mitrella, Chaetomitriopsis, Chaetomitrium, Chaetophora, Chamebrium, Humberlainia, Chameleon, Cheilotela, Henia, Chileobryon, Chionoloma, Chionostomum, Chorisodontium, Chryso-hypnum, Chrysoblastella, Chrysocladium, Chrysohypnum, Cinclidium, Circulifolium, Qi rriphyllum, Cladastomum, Cladomnion, Cladofascum , Cladopodanthus, Cladopodanthus, Claopodium, Clasmatodon, Clastobriella, Clastobryophilum, Clastobriopsis, Clastobrium, Claviteca, Cleistocarpidium, Cleistostoma, Climacium, Cnestrum, Codonoblepharon, Codonoblepharum, Codriophorus, Celidium, Coleochetium, Colobodontium, Conardia, Conomitrium , Conostomum, Coscinodon, Coscinodontella, Costesia K Tenidium, Ctenium, Kupressina, Curvicladium, Curviramea , Cyathophorella, Cyatoforum, Cyclodictyon, Cygniella, Cylicocarpus, Pigwort, Cynodoncilla, Cynodontium, Cynonthodium, Cirtohypnum, Cyrthomnium, Cyrthopodendron, Daltonia, Dasymitrium, Dawsonia, Dendrohypnum, Dendroalsia, Dendrocyatoforum, Dendrohypopterygium, Dendroli Gotrichum, Dermatodon, Desmatodon, Desmotheca , Dialitrichia, Diaphanophyllum, Dichelodontium, Dichelima, Dichodontium, Dicladiella, Diknemoloma, Dikranella, Dicranodon, Dicranodontium, Dicranodontium, Dicranoveusia, Dicranum, Didymodon, Dimerodontium, Dimorphocladon, Diobelon, Diobelonella, Difascum, Difiscium, Diplocomium, Diploneu ron, Diplostichum, Discelium, Discophyllum , Dissodon, Distichia, Distichium, Distichophyllidium, Distichophyllum, Ditrichopsis, Dietrichum, Dixonia, Dolichomithra, Dolichomitriopsis, Dolotortula, Donnellia, Donrichardsia, Dorcadion, Dosia, Drepanium, Drepano-hypnum, Drepanocladus, Drepanophyllaria, Drepanophyllum, Drum mondia, Dryptodon, Druzenia, Dutiella , Eccremidium, Echinodiopsis, Echinodium, Echinophyllum, Ectropotheciella, Ectropotheciopsis, Ectropothecium, Eleuthera, Elarvea, Elmeriobrium, Elodium, Encalyptus, Endotrichella, Endotrichellopsis, Endotrichum, Entodon, Entostodon, Entosthemenium, Eobruchia, Eochipopterygio psis, Aeoleukodon, Eosphagnum, Ephemerella, Ephemeridium, Ephemeropsis , Ephemerum, Epipterygium, Eremodon, Eriodon, Eriopus, Erpodium, Erythrobarbula, Erythrodontium, Erythrophyllastrum, Erythrophyllopsis, Erythrophyllum, Ezenbekia, Eucamptodontopsis, Eucatagonium, Eucladium, Evephemerum, Evmiurium, Euptychium, Eurynchiadelphus, Eurynchiastrum, Eurynchiella, Eurynchium, Eurogynum, Eustychia, Euzygodon , Exodiction, Exostratum, Exsertotheca, Fabrolesceia, Fabronialschirodon, Fabronidium, Fallaciella, Fauriella, Felipponea, Fiedleria, Phyphealsoteciadelphus, Fissidens, Flabellidium, Fleischerobrium, Floribundaria, Florschueziella, Flowersia, Fontinalis, Foroella, Forsstremia, Framiella , Funaria, Funariella, Gammiella, Gangvleia , Garkeya, Garovalgia, Gasterogrimmia, Hehebia, Gemmabrium, Georgia, Gertrudia, Gertrudiella, Gigaspermum, Giraldiella, Globulina, Globulinella, Glossadelfus, Glyphomitrium, Glyphomitrium, Glyfothecium, Glyptothecium, Gollania, Gongronia, Goniobrium, Goniomitrium, Gradstey niya, Grimmia, Grucilla, Guembelia , Guerramontesia, Gymnostomyella, Gymnostomum, Gyroveisia, Gabrodon, Gabrodonlshibealvatsukiella, Gageniella, Hematocaulis, Gampeella, Gampeohypnum, Gandeliobrium, Haplocladium, Haplodon, Haplodontium, Haplogimenium, Haptimenium, Harpidium, Harpophyllum, Garrisonia, Harveya, Hebantialtacillus, Hedenesia, Hedenasiastrum, Hedwigia, Hedwigidium , Helicoblepharum, Helicodontiadelphus, Helicodontium, Heliconema, Helicophyllum, Helodium, Hemiragis, Henicodium, Gennediella, Herpetineuron, Herzogiella, Heterocladium, Heterodon, Heterophyllium, Hildebrandzilla, Gilpertia, Hymantocladium, Goloblepharum, Holodontium, Holomitriopsis, Golom itrium, Gomalia, Gomaliadelphus, Gomaliodendron, Gomaliopsis , Homalothecyella, Homalothecium, Homomallium, Gondella, Hookeria, Hookeriopsis, Gorikavea, Gorridohypnum, Gusnocilla, Hyalophyllum, Hydrocryphealsodrepanium, Hydrogonium, Hydropogon, Hydropogonella, Hygroamblistegium, Hygrodicranum, Hygrohypnella, Hygrohypnum, Hylocomiadelphus , Hylocomiastrum, Hylocomiopsis, Hylocomium, Hymenodon, Hymenodontopsis, Hymenoloma , Hymenostomum, Hymenostiliella, Hymenostilium, Hyocomium, Hyophila, Hyophiladelphus, Hyophilopsis, Hypnella, Ginpitis, Hypnobartlettia, Hypnodendron, Hypnum, Hypodontium, Hypopterygium, Imbribrium, Indopottia, Indothuidium, Indusiella, Inouethuidium, Isopterygiopsis , Isopterygium, Isotheciopsis, Isothecium, Egerina, Egerinopsis , Jaffueliobrium, Yuratzkeella, Kieria, Kindbergia, Kingiobrium, Cleioveisiopsis, Coponenia, Kurohimehypnum, Lamprophyllum, Leersia, Leiodontium, Leiomela, Leiomitrium, Leiotheca, Lembophyllum, Lepidopylidium, Lepidopilum, Leptangium, Leptobarbula, Leptobrium, Leptokladium iella, Leptocladium, Leptodiccium, Leptodoncilla, Leptodontiopsis , Leptodontium, Leptohymenium, Leptofascum, Leptopteryginandrum, Leptostomopsis, Leptostomum, Leptotheca, Leptotrichella, Leptotrichum, Lepyrodon, Lepyrodontopsis, Leracia, Leracilla, Lescureia, Lesceia, Lesceadphus, Lesqueella, Lesceodon, Lesceodontopsis, Lesquereuxia, Leu cobrium, Leukodon, Leukodontella, Leukolepis, Leucoloma , Leukomium, Leukoperihetium, Leukofanella, Leukofanes, Levierella, Limbella, Limnobium, Limprichtia, Lindbergia, Lindigia, Lesceobrium, Leskipnum, Loisobrium, Luseria, Lophiodon, Lopidium, Lorencia, Lorenziella, Loxotis, Ludorugbia, Luisierella, Liellia, Macgregorella, Macuniella , Macrocoma . tus, Metadisticho

- 12 045836 phyllum, Metanekera, Meteoridium, Meteoriella, Meteoriopsis, Meteorite, Metzlerella, Metzleria, Mikralsopsis, Microbrium, Microcampylopus, Microcrossidium, Microktenidium, Microdus, Microeurynchium, Micromitrium, Micropoma, Microthamnium, Microteciella, Microtuidi mind, Micah, Milichhoferia, Mildea, Mildeella , Myronia, Mitrobrium, Mittenia, Mittenotamnium, Mittiridium, Miyabeya, Mniadelphus, Mniobrium, Mniodendron, Mniomalia, Mnium, Menkemeyer, Molendoa, Mollia, Morinia, Moseniella, Muelleriella, Muellerobrium, Muskoflorsvetsia, Muskoersogia, Myrinia, Miurella, Miuriopsi s, Miurium, Miuroclada , Nanobrium, Nanomitriopsis, Nanomitrium, Nekera, Nekeradelphus, Nekerites, Nekeropsis, Nematocladia, Neobarbella, Neocardotia, Neodicladiella, Neodolichomitra, Neoiophila, Neolescurea, Neolindbergia, Neomacunia, Neomeesia, Neonoguchia, Neophonics, Neorutenbergia, Neosarpi ella, Niphotrichum, Nobregea, Nogopteryum, Noguhiodendron , Notoligotrichum, Ochiobrium, Ochrobrium, Ochirea, Octodiceras, Oedicladium, Edipodiella, Oedipodium, Okamuraya, Oligotrichum, Oncoforsus, Oreas, Oreoveisia, Orontobrium, Orthoamblystegium, Orthodicranum, Orthodon, Orthodontium, Orthodontopsis, Orthogrimmia, Orthomitrium, Ortomnion, Ortomniopsis, Ortopus, Orthopixis . ella, Plustriella, Pankovia, Papillaria, Papillidiopsis, Paraleucobrium , Paramiurium, Pararacocarpus, Parisia, Pelecium, Pendulotecium, Pentastichella, Penzigiella, Peromnion, Pharomitrium, Fasconica, Fascopsis, Fascum, Filibercilla, Philonotis, Philophyllum, Photinophyllum, Phyllodon, Phyllodrepanium, Phyllogonium, Fiscomitrella, Fiscomitrium, Physedium, Picobri mind, Pictus, Piletium , Pilopogon, Pilopogonella, Piloseriopus, Pilotrichella, Pilotrichidium, Pilotrichum, Pinnatella, Piraeus, Pyreella, Plagiobrioides, Plagiobrium, Plagiomnium, Plagiopus, Plagioracelopus, Plagiothecium, Plasteurhynchium, Platydictia, Platygiriella, Platygirium, Platyhypnidium, Platyhypnum , Platiloma, Platilomella, Platinevron, Plavbelia . Porotamnium, Porotrichella, Porotrichodendron, Porotrichopsis , Porotrichum, Potamium, Pottia, Pottiopsis, Povellia, Povelliopsis, Pringleella, Prionidium, Prionodon, Pseudatrichum, Pseudefemerum, Pseudisothecium, Pseudoamblistegium, Pseudobarbella, Pseudobraunia, Pseudobrium, Pseudocalliergon, Pseudocampilium, Pseudochorizodontium, Pseudocrossidium, Pseudodimerodontium, Pseudodistichium, Pseudoditrichum, Pseudohygrohypnum, Pseudohyophila , Pseudohypnella, Pseudoleskeia, Pseudoleskeella, Pseudoleskeopsis, Pseudopylecium, Pseudopilotrichum, Pseudopleuropus, Pseudopoly, Pseudopterobrium, Pseudoracelopus, Pseudorhynchostegiella, Pseudoscleropodium, Pseudosymblepharis, Pseudotim Myella, Pseudotrismegistia, Psilopylum, Pteriginandrum, Pterobryella, Pterobryidium, Pterobryon, Pterobryopsis, Pterogoniadelphus, Pterogonidium, Pterogoniella, Pterogonium . , Pyramitrium, Pyromitrium, Pyrrhobrium, Questicula, Rakelopodopsis , Raquelopus, Racomitrium, Racopilum, Radulina, Raineria, Rauia, Rawella, Regmatodon, Reimersia, Remiella, Renauldia, Rhabdodontium, Rhabdoveisia, Racocarpus, Racopilopsis, Rhamphidium, Raphidorrhynchium, Raphidostegium, Raphidostichum, Rexophyllum, Rhizofabronia, Rhizogonium, Rhizoghi pnum, Rhizomnium, Rhizopelma , Rhodobrium, Rhyncho-hypnum, Rhynchostegiella, Rhynhostegiopsis, Rhynchostegium, Ristophyllum, Rytididelphus, Rytidiastrum, Rytidiopsis, Rytidium, Richardsiopsis, Rygodiadelphus, Roellia, Rosulabrium, Rottleria, Rutenbergia, Selania, Sagenotortula, Saintelenia, Saitoa, Site Obrium, Saitoella, Sanionia, Saproma , Sarkoneurum, Sarmentipnum, Sazaokea, Sauloma, Scabridens, Schimperella, Schimperobrium, Schistidium, Schistomitrium, Schistophyllum, Schistostega, Schizomitrium, Schizimenium, Schliefakea, Shloteymia, Schraderobrium, Schwetschkeya, Schwetshkeopsis, Sciadocladus, Sciar Omiella, Sciarmiopsis, Sciaromium, Sciurohypnum, Sclerodontium , Sclerogynum, Scleropodiopsis, Scleropodium, Scopelophila, Scorpidium, Scorpiurium, Sculeria, Scytalina, Sebillea, Senemobrium, Secra, Seligeria, Sematophyllites, Sematophyllum, Semibarbula, Serpoleskeia, Serpotortella, Sarpiella, Sevoskia, Sigmatella, Symophyllum, Simplicidens, Sinocalliergon, Sinskea, Scytophyllum , Scottsbergia, Solmsia, Solmsiella, Sorapilla, Spherangium, Spherocephalus, Spherothecium, Sphagnum, Spiridentopsis, Spirula, Splahnum, Sporledera, Spruceella, Squamidium, Stableria, Steerecleus, Stereobryon, Stegonia, Stellariomnium, Stenocarpidiopsis, Stenodesmus, St Enodictyon, Stenotheciopsis, Stenothecium, Steppomitra , Stereodon, Stereodontopsis, Stereohypnum, Steyermarkiella, Stokesiella, Stonea, Stoneobrium, Straminergon, Straminergon, Streblopilum, Streblotrichum, Streimannia, Strephedium, Streptocalypta, Streptocolea, Streptopogon, Streptotrichum, Stremia, Strombulidens, Strucia, Strucia, Stylo comium, Svartsia, Symblepharis, Symphiodon .

- 13 045836 elementium, Tetrodontium, Tamniella, Tamniopsis, Tamnium, Tamnobrium, Tamnomalia, Telia, Tiemea, Tuidiopsis, Tuidium, Thiridium, Tyzanomitrion, Thymmia, Timmiella, Timokoponenia, Toloxis, Tomentipnum, Tortella, Tortula, Tuvia, Tuviodendron, Trachybrium, Trachycarpidium , Trachycladiella, Trachycystis, Trachyloma, Trachymitrium, Trachiodontium, Trachyphyllum, Trachythecium, Trafixifium, Trematodum, Trichodon, Trichodontium, Tricholepis, Trihosteleum, Trichostomopsis, Trichostomum, Tridontium, Trigonodictyon, Tripterocladium, Triquetrella, Trismegistia, Tristichium, Tu Erkeimia, Uleastrum, Uleobrium, Ulota , Uncleyakia, Valdonia, Venturiella, Verrucidens, Vesicularia, Visekulariopsis, Vetiplanaxis, Viridivellus, Vittia, Voitia, Vroliheidia, Warburgiella, Vardia, Warnstorfia, Webera, Veisiodon, Veisiopsis, Veissia, Veissiodicranum, Werneriobrium, Weymouth ia, Viikia, Wildia, Villia, Wilsoniella , Yunnanobryon, Zelometeorium, Zygodon, Zygotrichia.

According to another preferred embodiment of the present invention, one, more or all of the above plants are at least one hornwort selected from the group consisting of the following genera: Antoceros, Dendroceros, Folioceros, Gattorioceros, Leiosporoceros, Megaceros, Mesoceros, Notoceros , Notothilas, Paraphymatoceros, Theoceros, Theomegaceros, Phymatoceros, Spherosporoceros.

A preferred embodiment of the present invention relates to the application described herein where the biocementation reaction is not an exothermic reaction. In the context of this text, an exothermic reaction is a reaction in which energy in the form of heat is released into the environment at constant pressure, in the preferred embodiment of the present invention - a reaction in which, at constant pressure, the temperature of the precursors, intermediates and/or products during biocementation reaction increases by more than 5°C, in a more preferred embodiment of the present invention - by more than 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100°C (relative to the initial temperature before the start of the biocementation reaction ).

Another aspect of the present invention relates to a method of preventing or reducing the growth of plants, in a preferred embodiment of the present invention, the growth of weeds, on/in a substrate, consisting or containing the following steps:

(a) identifying the substrate to be treated on/in which plant growth, in a preferred embodiment of the present invention the growth of weeds, is to be prevented or reduced, (b) providing a mixture (as defined above), (c) application and/or administration the mixture provided in step (b) onto/into the substrate to be treated in an amount sufficient to provide bio-cementation, and (d) the formation of a bio-cement (as defined above) layer so as to prevent or reduce the growth of plants or weeds on/ in the substrate.

According to a preferred embodiment of the method of the present invention, (only) application of the mixture provided in step (b) to/to the substrate to be treated occurs in step (c). According to another preferred embodiment of the method of the present invention, the application and subsequent introduction, for example by mixing, of the mixture provided in step (b) onto/into the substrate to be treated occurs in step (c).

According to another preferred embodiment of the method of the present invention, (only) the introduction of the mixture provided in step (b) onto/into the substrate to be treated occurs in step (c).

According to another preferred embodiment of the method of the present invention, the substrate or parts thereof identified in step (a) are removed from the original location, mixed with the mixture provided in step (b) in an amount sufficient to provide biocementation (for example, in a mixer ), the resulting mixture is returned to the original substrate location (or, in an alternative embodiment of the present invention, moved to another location where the biocement layer is to be formed), followed by step (d), as described herein. In such a preferred embodiment of the present invention, step (c) of the method described herein is omitted.

Depending on the form (solid or powder, liquid or gel or paste) of the mixture provided in step (b) of the method of the present invention (see the above explanation of this purpose), the application and/or administration in step (c) may occur differently. The powder mixtures may, for example, be dispersed throughout the substrate to be treated and/or incorporated into the substrate. Liquid mixtures, for example, are poured or sprayed onto the substrate to be treated, and are optionally subsequently incorporated into the substrate. In an alternative embodiment of the present invention, a single application and/or introduction of the mixture provided in step (b) onto/into the substrate to be treated is usually sufficient to form the biocement layer defined in step (d) of the method of the present invention. In a preferred embodiment of the present invention, a single application of the mixture provided in step (b) to/to the substrate to be treated is sufficient to form a biocement layer, specifically

- 14 045836 th at step (d) of the method according to the present invention.

Thus, one skilled in the art will understand that biocementation (as defined herein) is particularly effective in the method of the present invention at a certain application volume or a certain concentration of the mixture from step (b) (see also the preferred number of organism cells or organisms in mixture to be used according to the present invention, as defined above). According to our own research, the application volume of the mixture to be used according to the present invention (as defined above), in a preferred embodiment of the present invention, is at least 0.1 l/m ² , in a more preferred embodiment of the present invention, according to at least 0.5 l/m ² , in a more preferred embodiment of the present invention, at least 1.0 l/m ² , in a more preferred embodiment of the present invention, at least 2.0 l/m ² at least 3.0 l/m ² , at least 4.0 l/m ² or at least 5.0 l/m ² and/or in a more preferred embodiment of the present invention - at least at least 20.0 l/m ² , in a more preferred embodiment of the present invention at least 10.0 l/m ² .

For an efficient biocementation process in step (d) of the method of the present invention, it is an advantage of the present invention that the system of the mixture to be used in the present invention and the substrate (as defined herein) has a water content of more than 10% by weight calculated for the total mass of the specified system. If the mixture to be used according to the present invention is used in powder form (as defined above) in step (b) of the method of the present invention, and if the substrate in step (a) or (c) of the method of the present invention is also substantially free of water , so that the water content of said system is 10% by weight or less based on the total weight of the system, then the advantage of carrying out the method of the present invention is that the method comprises the further step of adding a sufficient amount of water or an aqueous solution to mixture from step (b) of the method before or after application or introduction onto/into the substrate to be treated, so that the water content of said system is more than 10% by weight based on the total weight of the results of said system. In an alternative or concurrent embodiment of the present invention, an appropriate amount of water or an aqueous solution may be added to the substrate to be treated before or after application or administration of the mixture provided in step (b) of the method of the present invention.

Moreover, if the method of the present invention is applied outdoors, then the advantage of the present invention is that the method is not carried out, for example, in the event of heavy rain or wind. Heavy rain or wind could potentially result in loss or significant dilution of the mixture to be used according to the present invention before the formation of the biocement layer (step (d)), which could prevent the formation of the biocement layer and/or negatively affect its strength and/or thickness. After applying or introducing the mixture provided in step (b) of the method of the present invention onto/into the substrate to be treated, that is, in step (d) of the method of the present invention, the formation of a biocement layer occurs, in a preferred embodiment of the present invention, during an incubation period of at least 6 hours, in a preferred embodiment of the present invention at least 24 hours, in a more preferred embodiment of the present invention at least 48 hours, during which, in a preferred embodiment of the present invention invention, there is no amount of rain, wind or artificial irrigation present that would result in significant loss of the mixture to be used according to the present invention. The required incubation period for the formation of the biocement layer in step (d) of the method of the present invention depends on various environmental parameters, such as indoor or outdoor temperature and humidity, as well as on the volume of mixture used. Considering the fact that during the specified incubation period of at least 6 hours, in the preferred embodiment of the present invention at least 24 hours, in the more preferred embodiment of the present invention at least 48 hours, rain or wind may cause significant losses of the mixture to be used according to the present invention, the advantage of the present invention is to repeat steps (b)-(d) of the method of the present invention the required number of times, in the preferred embodiment of the present invention one, two, three or more times, until until sufficient thickness and strength of the biocement layer is achieved to prevent or reduce the growth of plants, in the preferred embodiment of the present invention, weeds, on/in the substrate. In another or alternative embodiment of the present invention, the advantage of the present invention is to repeat steps (b)-(d) of the method of the present invention, in the preferred embodiment of the present invention one, two, three or more times if the thickness and/or strength of the biocement layer formed on/in the substrate decreases over time due to weathering and/or natural degradation and which is therefore no longer sufficient to prevent or reduce plant growth, in the preferred embodiment on

- 15 045836 worthwhile invention - growth of weeds, on/in the substrate.

The thickness of the biocement layer can be determined by manual measurement after mechanical destruction of the layer using a gauge clamp. In an alternative embodiment of the present invention, various (non-destructive) measurement methods used in construction, agriculture, geology or other applications (eg, MIT-SCAN-T2 handheld device) can be used depending on the thickening thickness. The thickness of the biocement layer includes a section of the substrate that thickens when the mixture is added.

The strength of the biocement layer corresponds to the destructive force (in Newtons (N)) that must be applied to destroy the biocement layer. The destruction of the biocement layer is the point at which (plastic) deformation of the layer no longer occurs as a result of the application of force, but rupture of the layer (biocementation) occurs. Failure is recognized by a decrease in the measured force. The breaking force (maximum force measurement value) can be determined by the following method: This method is based on the standardized method for testing the strength of cement DIN EN 196-1:2005-05. According to the manufacturer, breaking force is measured using a digital (breaking) force measuring instrument. The test specimen is pressed into the sample (until failure) using a crank test bench and the force applied is continuously measured. The average breaking force is calculated from several measurements (>3). The average breaking force, in a preferred embodiment of the present invention, is from 0.5 to 1000 N, in a more preferred embodiment of the present invention, from 3 to 300 N.

The method described herein is also preferred, wherein the formed biocement layer (in step (d)) of the method of the present invention) has a permeability coefficient (water) in the range of more than 10 ^-9 to 100 m/s, preferably implementation of the present invention - from more than 10 ^-9 to 10 ^-3 m/s, in another preferred embodiment of the present invention - from more than 10 ^-8 to 10 ^-3 m/s.

Optionally, after step (d) of the method of the present invention, a further step (e) may take place, which comprises or consists of monitoring whether plant growth, in a preferred embodiment of the present invention, weed growth, is prevented or reduced. This control can be carried out, for example, by determining the rate of growth of plants or weeds by manual visual assessment, as described in the following examples. Step (e) of the method of the present invention can be repeated at regular intervals if necessary, for example every 24 or 48 hours.

The method described above is preferred, wherein the substrate is selected from the group consisting of sand, soil, in the preferred embodiment of the present invention - ground soil and plant soil, humus, crushed stone, gravel, clay, silt, sawdust, paper, cardboard , particle boards, softwood, limestone, coal and its mixtures.

Another preferred substrate used in the method of the present invention is selected from the group consisting of organic and inorganic material and mixtures thereof in which plant growth is possible, in a preferred embodiment of the present invention heavy mineral sand, fine sand, natural sand, quartz sand, crystalline quartz sand, bird feeding sand, gravel sand, cohesive sand, crushed sand, quartz flour, mineral mixture (stone, stone chips, gravel), triple-fired sand, Savonier stone flour, plaster, loess, turf, limestone crushed sand, limestone flour, calcium carbonate (including polymorphs, derivatives and mixtures, as well as natural-based GCC (fine calcium carbonate - GCC) and synthetic PCC (precipitated calcium carbonate - PCC)), talc, dolomite, white lime (hydrate) , route, cements and mixtures, microsilica, chalk (mixture), marble, perlite, overburden, dump material, hematite, red chalk, magnesite, iron ore, steatite, soapstone, kaolin, marl, alumina, attapulgite, clay minerals, bentonite, zeolite, (calco)plaster, gravel, glass powder, aluminum oxide, aluminum hydroxide, magnesium oxide, calcium oxide, calcium hydroxide, magnesite, slate powder, pumice, cristobalite (sand), Roman cement, bauxite, pyrites, sphalerites, silicates, oxides, carbonates, wood (chips), mulch, alluvial soil, laterite, red iron ore, ash, (wood ash, fly ash, bone ash), soils from (pig) farms, standard LUFA soils (see. eg http://www.lufaspeyer.de) or mixtures thereof.

The method described above is also preferred, wherein the substrate is a plot of land or crop area, in the preferred embodiment of the present invention - in the air, such as, for example, a garden plot, an adjacent area of terraces or entrances and exits, a cultivated plot, a fruit garden a garden, vineyard, tree nursery, park, part of developed land or urban area, road, street, footpath, railway line or area used for industrial production.

Depending on the properties of the substrate to be treated, it may be advisable to add at least one of the above additives to the substrate (or component (i), (ii) and/or (iii) of the mixture provided in step (b)), e.g. , to improve the reactivity of the substrate with

- 16 045836 biocement formed during the method according to the present invention. The advantage here is that this results in a particularly hard or stable biocement layer, which is particularly effective at suppressing weed growth.

The method of the present invention allows, for example, to seal and/or cure the joint surfaces of terraces, entrances and exits, driveways, roads or walkways, or open areas using biocementation, thereby effectively suppressing plant growth, in a preferred embodiment of the present invention weeds in/on these substrates. It is also possible to use the method of the present invention for the suppression of weeds in agriculture, for example, on agricultural land used for growing cereals or fruits.

Therefore, a preferred embodiment of the present invention relates to the method of the present invention described herein, wherein the biocement layer formed in step (d) allows crop plants to grow but prevents or reduces the growth of new weeds.

A preferred embodiment of the present invention is the method described above, wherein the plant or weed is selected from the group consisting of dicotyledonous plants of the following genera: Abutilon, Gooseberry, Kokorysh, Shchiritsa, Ambrosia, Ankhusa, Anagallis, Anode, Anthemis, Nevzrachnitsa, Arabidopsis, Quinoa , Crescent, Daisy, Seed, Sverbiga, Shepherd's purse, Thistle, Cassia, Cornflower, Goosefoot, Chrysanthemum, Thistle, Hemlock, Conise, Consolidum, Bindweed, Datura, Descurainia, Desmodium, Emex, Equisetum, Erigeron, Erodium, Erizimum, Euphorbia, Fumaria, Galeopsis, Galinzoga, Galium, Geranium, Hogweed, Hibiscus, Morning Glory, Kochia, Lathyrus, Warthog, Lathyrus, Lepidium, Lithospermum, Linaria, Lindernia, Crooked Flower, Mallow, Matricaria, Mint, Woodweed, Mollugo, Forget-me-not, Papaver, Farbitis, Plantain, Polygonum, Purslane, Ranunculus, Radish, Zherushnik, Rotala, Sorrel, Solyanka, Rasogus, Sesbania, Sida, Sinapis, Gooseberry, Solanum, Sow thistle, Sphenoclea, Stachys, Chickweed, Dandelion, Jarutka, Clover, Coltsfoot, Nettle, Veronica, Viola, Cocklebur; dicotyledonous plants of the following genera: Peanut, Beetroot, Brassica, Cucumis, Pumpkin, Helianthus, Carrot, Wisteria, Cotton, Morning Glory, Lettuce, Flax, Tomato, Tobacco, Beans, Peas, Solanum, Pea monocotyledonous plants of the following genera: Aegilops, Zhitnyak, Bentgrass , Alopecurus, Metlitsa, Oats, Brachiaria, Bonfire, Cenchrus, Commelina, Pigwort, Cyperus, Dactyloctenium, Crabgrass, Barnyardgrass, Eleocharis, Eleusina, Eragrostis, Woolgrass, Fescue, Fimbristylis, Heteranthera, Imperata, Murraya, Juncus, Leptochloa, Tares, Monochoria , Panicum, Paspalum, Canary grass, Timothy grass, Bluegrass, Rothbellia, Sagittaria, Scirpus, Setaria, Sorghum; and monocotyledonous plants of the following genera: Allium, Pineapple, Asparagus, Oats, Barley, Rice, Panicum, sugar cane, Rye, Sorghum, Triticale, Wheat, Corn; mosses of the class liverworts (Marshantiaceae), hornworts (Anthocerotaceae), mosses (Bryopsidae);

According to a preferred embodiment of the method of the present invention, one, more or all of the above plants are at least one liverwort selected from the group consisting of the following genera: Acolea, Acrobolbus, Acrochilla, Acromastigum, Acroscyfella, Acroscyphus, Acrostolia, Adelocolia , Aichisonella, Alicularia, Allisonia, Allisonella, Alobiella, Alobiellopsis, Amazoopsis, Amphicephalosia, Amphilophocolea, Andrewsianthus, Aneura, Anomacaulis, Anomoklada, Anomilia, Antelia, Antelis, Aphanolezhenea, Aplosia, Apomarsupella, Apometgeria, Apotreubia, Arachniopsi s, Arctosciphus, Arnellia, Ascidiota . Ephalolobus, Cephalomitrion, Cephalosia, Cephalosiella, Cephalosiopsis , Keratolezhenea, Caesius, Chaetophyllopsis, Chiastokavlon, Chiloscyphus, Chloranthelia, Honecolea, Cladomastigum, Cladopodiella, Clandarium, Clasmatocolea, Cololezhenea, Kolura, Conocephalum, Conoscyphus, Corsinia, Kronizia, Crossogyna, Cryptochila, Cryptocolea, Cryptocolepsis, Cryptomithrium, Cryptostipula, Cryptothallus, Cuspidatula , Cyanolophocolea, Cyathodium, Cylindrocolea, Delavayella, Dendrobazzania, Dendromastigophora, Denotarysia, Dichiton, Dinkleria, Diplocolea, Diplophyllum, Duinia, Drepanolezhenea, Drucella, Dumortiera, Dumortieropsis, Enigmella, Eocalypogeia, Eoisotachis, Eopleurosia, Eotry hokolea, Eremonotus, Eucalyx, Evansia, Evansianthus . Haplomitrium, Harpalezhenea, Garpanthus, Gattoria , Gattorianthus, Gattoriella, Hepatostolonophora, Herbertus, Herpetium, Herpocladium, Herzogianthus, Herzogobrium, Heterogemma, Heteroscyphus, Horikavella, Hyalolepidosia, Hygrobiellalvatsukia, Hygrolembidium, Hygrophila, Hymenophyton, Hypoisotachis, Isolembidium, Isotakis, Jamsoni ella, Jensenia, Yubula, Yubulopsis, Jungermannia, Jungermannites , Crunodiplophyllum, Curzia, Cymatocalyx, Lamellocolea, Leiocolea, Leiomithra, Leiomilia, Leioscyphus, Lezhenea, Lembidium, Lepidogyna, Lepidolena, Lepidosia, Leptoleugea, Leptophyllopsis, Leptoscyphopsis, Leptoscyphus, Letokolea, Liochlena, Lobatiriccardia, Lofo rut, Lophonardia, Lofosia, Lofosiopsis, Lunularia , Macrodiplophyllum, Maculia, Mackinoa, Mannia, Marchantia, Marchesinia, Marsupella, Marsupidium, Massula, Massularia, Mastigobrium, Mastigopelma,

- 17 045836

Mastigophora, Mastigopsis, Mesoptychia, Metacalipogea, Metaigrobiella, Mezgeria, Mezgeriopsis, Micrizophylla, Microlezhenea, Microlepidosis, Micropterygium, Misutania, Mnioloma, Murkia, Monocarpus, Monoclea, Monodactylopsis, Monosolenium, Mytilopsis, Nanomarsupella, Nardia, Neesioscyphus, Neogrollea, Neochodzonia, Neotrichocolea, Noteroclada, Notogymnomitrion, Notostrepta, Notscyphus, Novellia, Obtusifolium, Odontolezhenea, Odontoschism, Oleolophosia, Oximitra, Pachyglossa, Pachyschistochila, Pallavicinia, Paracromastigum, Paraschistochila, Patarola, Pediophyllopsis, Pediophyllum, Pellia, Peltolepsis, Perdusenia, Perssoniella, Petalophyllum, Phycolepidosis, Phyllotallia, Physiothium, Physoteca, Pisanoia, Plagiochasma, Plagiochilla, Plagiochilidium, Plagiochilion, Platicaulis, Plectocolea, Pleurante, Pleuroclada, Pleurocladopsis, Pleurocladula, Pleurosia, Podante, Podomitrium, Porella, Prasanthus, Preissia, Prionolobus, Protolophosia, Protomarsupel la, Protosysgiella, Protosyzygiella, Pseedocephalosia, Pseedocephalosiella, Pseudolofocolea, Pseudolophosia, Pseudomarsupidium, Pseudoneura, Pseudotritomaria, Psiloclada, Pteropsiella, Ptilidium, Radula, Rebulia, Rhizocaulia, Rhodoplagiochyla, Riccardia, Riccia, Ricciella, Ricciocarpos, Riella, Roivainenia, Ruisanthus, Ruth tnerella, Saccobasis, Saccogina, Sandeothallus, Sarkotsifos, Sarcomitrium, Sauteria, Scapania, Scaphophyllum, Schiffneria, Schisma, Schistochila, Schistochylaster, Schistochilopsis, Scofildia, Sendtner, Seppelzia, Sevardiella, Simodon, Solenostoma, Sothbia, Spherocarpus, Sphagnoecetis, Sprucella, Steereella, Stereocolea, Stenorri pis, Stephandium, Stephiniella, Stefaniellidium, Stephensoniella, Symphyogyna, Symphyogynopsis, Symphyomitra, Synchimenium, Sysgiella, Teniolezhenea, Thargyonia, Tegulifolium, Telaranea, Tallocarpus, Treubia, Triandrophyllum, Trichocolea, Trichocoleopsis, Trichostilium, Trichotemnoma, Trilophosia, Tritomaria, Tilimanthus, Vanaea, Vandiemenia , Verdornia, Vetaforma, Vettsteinia, Viesnerella, Xenochila, Xenothallus, Zoopsidella, Zoopsis.

According to another preferred embodiment of the method of the present invention, one, more or all of the above-mentioned plants are at least one moss selected from the group consisting of the following genera: Abietinella, Acanthocladiella, Acanthocladium, Acanthodium, Acanthorhynchium, Akavlon, Akavlonopsis, Achrophyllum, Acidodontium, Acrocladium, Acroporium, Akroschisma, Actinodontium, Actinotuidium, Adelothecium, Equatorella, Erobryidium, Erobryopsis, Erobryum, Erolindigia, Algaria, Aligrimmia, Alleniella, Allioniellopsis, Aloina, Aloinella, Alofosia, Alzia, Amblyodon, Amblyodum, Amblystegiella, Amblystegium, Amblytropis, Ambukhanania, Amphidium, Amphoridium, Amphoritheca, Anakalipta, Anakamptodon, Anacolia, Ancistrodes, Andoya, Andreaea, Andreobrium, Aniktangium, Anisothecium, Anodon, Anodontium, Anoectangium, Anomobrium, Anomodon, Antitrichia, Ongstremia, Ongstremiopsis, Apalodium, Aphanorrhegma, Apiocarpa, Aplodon, Apterygium, Aptychella, Aptychopsis, Aptychus, Arbuscula, Arbusculopopterygium, Archaephemeropsis, Archidium, Arctoa, Argyrobrium, Arthrocormus, Aschisma, Aschistodon, Asteriscium, Astomyopsis, Astomum, Astrodontium, Astrophylum, Atractylocarpus, Atrichopsis, Atrichum, Avlacomytri mind, Avlakomnium, Avlakopilum, Avstinella, Austrohondella, Austrophiliberciella, Baldwiniella, Barbella, Barberlopsis, Barbula, Bartramia, Bartramiopsis, Beveria, Bellibarbula, Benitotania, Bestia, Bissetia, Blindia, Boulaya, Brahelima, Brachydontium, Brachymenium, Brachymitrion, Brachiodus, Brachysteleum, Brachytheciastrum, Brachytecia ella, Brachythecium, Brachytrichum, Braithwaitea, Brunfelsia, Braunia, Breidleria, Breutelia, Brotera, Broterella, Broterobrium, Bruchia, Brychnia, Brimela, Bryoandersonia, Bryobeckettia, Bryobrittonia, Bryobrotera, Bryoceutospora, Bryochenea, Bryokrumia, Bryodixonia, Bryodusenia, Bryoerythrophyllum, Bryohaplocladium, Bryog Umbertia, Briomaltea, Briomaginia, Bryomnium, Bryonoguchia, Bryonorrhizia, Bryopyxia, Bryozedgvikia, Bryostreymannia, Bryotestua, Bryum, Bukiella, Buklandiella, Burnettiia, Buxbaumia, Callialaria, Callicladium, Callicosta, Callicostella, Callicostellopsis, Calliergidium, Calliergon, Kaloghypnum, Kalimperastrum, Kalimpers, Kalimperidium, Kalimperopsis, Calyptopogon, Calyptothecium, Calyptrochaete, Camptochaete, Camptodontium, Camptothecium, Campyadelphus, Campylidium, Campilium, Campylodontium, Campiphyllum, Campylopodiella, Campylopodium, Campylopus, Campylostelium, Canalogopterygium, Cardocia, Cardocilla, Caribeohypnum, Catagoniopsis, Catagonium, Catarinaea, Catarinella, Catharomnion, Katoskopium, Cecalifum, Ceratodom, Ceutospora, Ceutoteca, Chaetomitrella, Chaetomitriopsis, Chaetomitrium, Chaetophora, Chamebrium, Humberlainia, Chameleon, Cheilotela, Henia, Chileobryon, Chionoloma, Chionostomum, Chorizodontium, Chryso-hypnum, Chrysoblastella, Chrysocladium, Chrysohypnum, Cinclidium, Circulifolium, Cirriphyllum, Cladastomum, Cladomnion, Cladofascum, Cladopodanthus, Cladopodanthus, Claopodium, Clasmatodon, Clastobriella, Clastobryophilum, Clastobriopsis, Clastobrium, Clavitheca, Cleistocarpidium, Cleistostoma, Climacium, Cnestrum, Codonoblepharon, Codonoblepharum, Codriophorus, Celidium, Coleochetium, Colobodontium, Conardia, Conomitrium, Conostomum, Coscinodon, Coscinodontella, Costesia, Craspedophyllum, Cratoneurella, Cratoneuron, Cratoneuropsis, Crosbia, Crossidium, Crossomitrium, Crumia, Crumuscus, Cryphaea, Crifeadelphus, Cryptocarpon, Cryptodicranum, Cryptogonium, Cryptoleptodon, Cryptopapillaria, Cryptopodium, Cryptopodium, Cryptoteca, Cten Idiadelphus, Ctenidium, Ctenium, Kupressina, Curvicladium, Curviramea, Cyatophorella, Cyathoforum, Cyclodictyon, Cygniella, Cylicocarpus, Pigwort, Cynodoncilla, Cynodontium, Cynontodium, Cirto-hypnum, Cyrthomnium, Cyrtopodendron,

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Daltonia, Dasimitrium, Dawsonia, Dendrohypnum, Dendroalsia, Dendrocyatoforum, Dendrohypopterygium, Dendroligotrichum, Dermatodon, Desmatodon, Desmotheca, Dialitrichia, Diaphanophyllum, Dichelodontium, Dihelima, Dichodontium, Dicladiella, Diknemoloma, Dicranella, Dicranodon, Dicranodontsi mind, Dicranoysia, Dicranoeysia, Dicranum, Didymodon, Dimerodontium, Dimorphocladon, Diobelon, Diobelonella, Diphascum, Diphyscium, Diplocomium, Diploneuron, Diplostichum, Discelium, Discophyllum, Dissodon, Distichia, Distichium, Distichophyllidium, Distichophyllum, Ditrichopsis, Dietrichum, Dixonia, Dolichomithra, Dolichomi Triopsis, Chisel, Donnellia, Donrichardsia, Dorcadion, Dosia, Drepanium, Drepano-hypnum, Drepanocladus, Drepanophyllaria, Drepanophyllum, Drummondia, Dryptodon, Druzenia, Dutiella, Eccremidium, Echinodiopsis, Echinodium, Echinophyllum, Ectropotheciella, Ectropotheciopsis, Ectropothecium, Eleuthera, Elarvea, Elmer Iobrium, Elodium, Encalyptus, Endotrichella, Endotrichellopsis, Endotrichum, Entodon, Entostodon, Entostimenium, Eobruchia, Eohypopterygiopsis, Eoleucodon, Eosphagnum, Ephemerella, Ephemeridium, Ephemeropsis, Ephemerum, Epipterygium, Eremodon, Eriodon, Eriopus, Erpodium, Erythrobarbula, Erythrodontium, Er itrophyllastrum, Erythrophyllopsis, Erythrophyllum, Esenbekia, Eucamptodontopsis, Eucatagonium, Eucladium, Evephemerum, Eumyurium, Euptychium, Eurynchiadelphus, Eurynchiastrum, Eurynchiella, Eurynchium, Eurogynum, Eustychia, Euzygodon, Exodiction, Exostratum, Exsertotheca, Fabrolesceia, Fabronialschirodon, Fabronidium, Fallaciella, Fauriella, Felipponea, Fiedleria, Phifealsoteciadelphus, Fissidens, Flabellidium, Fleischerobrium, Floribundaria, Florschueziella, Flowersia, Fontinalis, Foroella, Forsstremia, Framiella, Funaria, Funariella, Gammiella, Gangvleia, Garkea, Garowalgia, Gasterogrimmia, Hehebia, Gemmabrium, Georgia, Gertrudia, Gertrudiella, Gigaspermum, Giraldiella, Globulina, Glob ulinella, Glossadelphus, Glyfomitrium, Glyphomitrium, Glyfothecium, Glyptothecium, Gollania, Gongronia, Goniobrium, Goniomitrium, Gradsteinia, Grimmia, Grucilla, Guembelia, Guerramontesia, Gymnostomyella, Gymnostomum, Gyroveisia, Gabrodon, Gabrodonlshibealvatsukiella, Gageniella, Hematocaulis, Gampeella , Gampeohypnum, Gandeliobrium, Haplocladium, Haplodon, Haplodontium, Haplogimenium, Haptimenium, Harpidium, Harpophyllum, Garrisonia, Harveya, Hebantialtacillus, Hedenesia, Hedenasiastrum, Hedvigia, Hedwigidium, Helicoblepharum, Helicodontiadelphus, Helicodontium, Heliconema, Helicophyllum, Gelodium, Hemiragis, Genicodium, Gennediella, Herpetineuron, Herzogiella, Heterocladium, Heterodon, Heterophyllium, Hildebrandcilla, Gilpertia, Hymantocladium, Holoblepharum, Holodontium, Holomitriopsis, Holomitrium, Gomalia, Gomaliadelphus, Gomaliodendron, Homaliopsis, Gomalothecyella, Gomalothecium, Homomallium, Gondella, Hookeria, Hookeriopsis, Gorikavea, Gorridogypnum, Gusno Cilla, Hyalophyllum, Hydrocryphealsodrepanium, Hydrogonium, Hydropogon, Hydropogonella, Hygroamblystegium, Hygrodicranum, Hygrohypnella, Hygrohypnum, Hylocomiadelphus, Hylocomiastrum, Hylocomiopsis, Hylocomium, Hymenodon, Hymenodontopsis, Hymenoloma, Hymenostomum, Hymenostiliella, Hymenostilium, Hyocomium, Hyophila, Hyophiladelphus, Hyophilopsis, Gi pnella, Ginpitis, Hypnobartlettia, Hypnodendron, Hypnum, Hypodontium, Hypopterygium, Imbribrium, Indopottia, Indotuidium, Indusiella, Inouethuidium, Isopterygiopsis, Isopterygium, Isotheciopsis, Isothetium, Egerina, Egerinopsis, Jaffueliobrium, Yuratzkeella, Kieria, Kindbergia, Kingiobrium, Cleioveisiopsis, Koponenia, Kurohimehypnum , Lamprophyllum, Leersia, Leiodontium, Leiomela, Leiomitrium, Leiotheca, Lembophyllum, Lepidopylidium, Lepidopilum, Leptangium, Leptobarbula, Leptobrium, Leptocladiella, Leptocladium, Leptodiccium, Leptodoncilla, Leptodontiopsis, Leptodontium, Leptohymenium, Leptofascum, Leptopteryginandrum, Leptostomopsis, Leptostomum, Leptotheca , Leptotrichella, Leptotrichum, Lepirodon, Lepirodontopsis, Leracia, Leracilla, Lescurea, Lesceia, Lesqueadphus, Lesqueella, Lesceodon, Lesceodontopsis, Lesquereuxia, Leukobrium, Leukodon, Leukodontella, Leukolepis, Leukoloma, Leukomium, Leukoperihetium, Leukofanella, Leukofanes, Levierella, Limbella, Limnobium, Limprichtia, Lindbergia, Lindigia, Leskeobrium, Leskipnum, Loisobrium, Luseria, Lophiodon, Lopidium, Lorencia, Lorenziella, Loxotis, Ludorugbia, Luisierella, Liellia, Macgregorella, Macuniella, Macrocoma, Macrodictium, Macroimenium, Macromitrium, Macrosporiella, Macrotamniella, Macrotamnium, Mamillariella, Mandoniella, Maschalanthus, Maschalocarpus, Mastopoma, Matteria , Meesia, Meiotheciella, Meiotheciopsis, Meiothecium, Meiotrichum, Mercea, Merceiopsis, Mesochete, Mesonodon, Mesotus, Metadistichophyllum, Metanequera, Meteoridium, Meteoriella, Meteoriopsis, Meteorum, Metzlerella, Metzleria, Mikralsopsis, Microbrium, Microcampylopus, Microcrossidium, Microkten idium, Microdus, Microeurynchium, Micromitrium, Micropoma, Microthamnium, Microteciella, Microtuidium, Micahea, Milichhoferia, Mildea, Mildeella, Mironia, Mitrobrium, Myttenia, Mittenotamnium, Mittiridium, Miyabeya, Mniadelphus, Mniobrium, Mniodendron, Mniomalia, Mnium, Menkemeyer, Molendoa, Mollia, Morinia, Moseniella, Muelleriella, Muellerobrium,

Muskoflorsvetsia, Muskoersogia, Myrinia, Miurella, Miuriopsis, Miurium, Miuroclada, Nanobrium, Nanomitriopsis, Nanomitrium, Nekera, Nekeradelphus, Nekerites, Nekeropsis, Nematocladia, Neobarbella, Neocardocia, Neodicladiella, Neodolichomithra, Neoiophila, Neolescurea, Neolindbergia, Neomacunia, Neomeesia, Neonoguchia, Neofonix, Neorutenbergia, Neosarpiella, Niphotrichum, Nobregea, Nogopterium, Noguhiodendron, Notoligotrichum, Ochiobrium, Ochrobrium, Ochirea, Octodiceras, Oedicladium, Oedipodiella, Oedipodium, Okamuraya, Oligotrichum, Oncoforsus, Oreas, Oreoveisia,

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Orontobrium, Orthoamblystegium, Orthodicranum, Orthodon, Orthodontium, Orthodontopsis, Orthogrimmia, Orthomitrium, Ortomnion, Ortomniopsis, Ortopus, Orthopixis, Orthorhynhidium, Orthorhynchium, Ortostichella, Ortostichidium, Ortostichopsis, Orthotheciella, Orthothecium, Orthothecium, Or totuidium, Orthotrichum, Osterwaldiella, Oticodium, Oxirrhynchium, Oxystegus, Pachyneuropsis, Pachyneurum, Paleocampylopus, Palamocladium, Palisadula, Paludella, Plustriella, Pankovia, Papillaria, Papillidiopsis, Paraleucobrium, Paramiurium, Pararacocarpus, Parisia, Pelecium, Pendulotecium, Pentastichella, Penzigiella, Peromnion, Pharomitrium, Fasco nika, Fascopsis, Fascum, Filibercilla, Philonotis, Philophyllum, Photinophyllum, Phyllodon, Phyllodrepanium, Phyllogonium, Fiscomitrella, Fiscomitrium, Physedium, Picobrium, Pictus, Piletium, Pilopogon, Pilopogonella, Piloseriopus, Pilotrichella, Pilotrichidium, Pilotrichum, Pinnatella, Piraeus, Pireella, Plagiobrioides, Plagiobrium, Plagiomnium, Plagiopus, Plagioracelopus, Plagiothecium, Plasteurhynchium, Platydicty, Platygiriella, Platygirium, Platyhypnidium, Platyhypnum, Platyloma, Platilomella, Platinevron, Plavbelia, Pleuriditrichum, Pleuridium, Pleurochaete, Pleurofascum, Pleuropus, Pleurortotrichum, Pleuroveisia, Pleurosium, P Leurozygodon, Poxiella, Podperea, Pecilophyllum, Pogonatum, Polya, Polla, Polymerodon, Polypodiopsis, Polytrichadelphus, Polytrichastum, Polytrichites, Polytrichum, Porotamnium, Porotrichella, Porotrichodendron, Porotrichopsis, Porotrichum, Potamium, Pottia, Pottiopsis, Powellia, Povelliopsis, Pringleella, Prionidium, Prionodon, Pseudatrichum, Ps eudafemerum, Pseudisothecium, Pseudoamblystegium, Pseudobarbella, Pseudobraunia, Pseudobrium, Pseudocalliergon, Pseudocampilium, Pseudochorisodontium, Pseudocrossidium, Pseudodimerodontium, Pseudodistichium, Pseudoditrichum, Pseudohygrohypnum, Pseudohyophila, Pseudohypnella, Pseudolesceia, Pseudoleskeella, Pseudoleskeopsis, Pseudopylecium, Pseudopilotrichum, Pseudopleuropus, Pseudopodia, Pseudopterobrium, Pseudoracelopus, Pseudorhynchosteghiella, Pseudoscleropodium, Pseudosymblepharis, Pseudothymmiella, Pseudotrismegistia, Psilopylum, Pteriginandrum, Pterobryella, Pterobryidium, Pterobryon, Pterobryopsis, Pterogoniadelphus, Pteragonidium, Pterogoniella, Pterogonium, Pterogoneurum, Pterogophyllum, Ptylium, Ptychodium, Ptychomitriopsis, Ptychomitri um, Ptychomniella, Ptychomnion, Ptychostomum, Puiggaria, Puiggariella, Puiggariopsis, Pulchrinodus, Pungentella, Purcellia, Pilesia, Pilesiadelpha, Pilesiella, Pilesiobrium, Pyramidula, Pyramitrium, Pyromitrium, Pyrrhobrium, Questicula, Rachelopodopsis, Rakelopus, Racomitrium, Racopilum, Radulina, Raineria, Rauia, Rauella, Regmatodon, Reimersia, Remieella, Renauldia, Rhabdodont ium, Rabdoweisia, Racocarpus, Racopylopsis, Rhamphidium, Raphidorrhynchium, Raphidostegium, Raphidostichum, Rexophyllum, Rhizofabronia, Rhizogonium, Rhizohypnum, Rhizomnium, Rhizopelma, Rhodobrium, Rhynchohypnum, Rhynchostegiella, Rhynhostegiopsis, Rhynchostegium, Ristophyllum, Rhytididelphus, Ritidiastrum, Ritidiopsis, Ritidium, Richardsiopsis, Rigodiadelphus, Roellia, Rosalabrium, Rottleria, Rutenbergia, Selania, Sagenotortula, Saintelenia, Saitoa, Saitobrium, Saitoella, Sanionia, Saproma, Sarconevrum, Sarmentipnum, Sazaokea, Sauloma, Scabridens, Schimperella, Schimperobrium, Schistidium, Schistomitrium, Schistophyllum, Schistostega, Schizomitrium, Schizimenium, Schliefakea, Shloteymia, Schraderobrium, Shwetschkeya, Shwetshkeopsis, Sciadocladus, Sciaromiella, Sciarmiopsis, Sciaromium, Sciuro-hypnum, Sclerodontium, Sclerogypnum, Scleropodiopsis, Scleropodium, Scopelophila, Scorpidium, Scorpiurium, Sculeria, Scytalina, Sebillea, Sen Hemobrium, Sekra, Seligeria, Sematophyllites, Sematophyllum, Semibarbula, Serpoleskeia, Serpotortella, Sarpiella, Sevoskia, Sigmatella, Simophyllum, Simplicidens, Sinocalliergon, Sinskea, Scytophyllum, Scottsbergia, Solmsia, Solmsiella, Sorapilla, Spherangium, Spherocephalus, Spherothecium, Sphagnum, Spiridentopsis, Spirula, Splaxnum, Sporle dera, Spruceella, Squamidium, Stableria, Sterecleus, Stereobryon, Stegonia, Stellariomnium, Stenocarpidiopsis, Stenodesmus, Stenodictyon, Stenotheciopsis, Stenothecium, Steppomitra, Stereodon, Stereodontopsis,

Stereohypnum, Steyermarkiella, Stokesiella, Stonea, Stoneobrium, Straminergon, Straminergon, Streblopilum, Streblotrichum, Streimannia, Strephedium, Streptocalyptus, Streptocolea, Streptopogon, Streptotrichum, Stremia, Strombulidens, Strucia, Strucia, Stylocomium, Swarcia, Symblepharis, Sim Phiodon, Symphisodon, Symphisodontella, Syntrichia, Sirropodon, Systegium, Taiwanobrium, Takakia, Tamariscella, Taxicaulis, Taxiphyllum, Taxitelyum, Tayloria, Teichodontium, Teniolophora, Teretidens, Terrestria, Tetracoscinodon, Tetraphidopsis, Tetraphis, Tetraplodont, Tetrapterum, Tetrastichium, Tetrodontium, Tamniella, Tamnio psis, Tamnium, Tamnobrium, Tamnomalia, Telia, Thiemea, Tuidiopsis, Tuidium, Thyridium, Tyzanomitryon, Thymmia, Timmyella, Thymokoponenia, Toloxis, Tomentipnum, Tortella, Tortula, Thuvia, Tuviodendron, Trachybrium, Trachycarpidium, Trachycladiella, Trachycystis, Trachyloma, Trachymitrium, Trachyodontium, Trachyphyllum , Trachythecium, Trafixifium, Trematodum, Trichodon, Trichodontium, Tricholepis, Trichosteleum, Trichostomopsis, Trichostomum, Tridontium, Trigonodictyon, Tripterocladium, Triquetrella, Trismegistia, Tristichium, Tuerkeymia, Uleastrum, Uleobrium, Ulota, Uncleyakia, Valdonia, Venturiella, Verruzi dens, Vesicularia, Visekulariopsis, Vetiplanaxis, Viridivellus, Vittia, Voitia, Vroliheidia, Warburgiella, Vardia, Warnstorfia, Webera, Veisiodon, Veisiopsis, Veissia, Veissiodicranum, Werneriobrium, Veimoutia, Viikia, Wildia, Willia, Wilsoniella, Yunnanobryon, Zelometeorium, Zygodon, Zygotrichia.

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According to another preferred embodiment of the method of the present invention, one, more or all of the above plants are at least one hornwort selected from the group consisting of the following genera: Antoceros, Dendroceros, Folioceros, Gattorioceros, Leiosporoceros, Megaceros, Mesoceros, Notoceros, Notothilas, Paraphymatoceros, Theoceros, Theomegaceros, Phymatoceros, Spherosporoceros.

Another preferred embodiment of the present invention is the method described above, wherein the mixture is present in liquid, gel, paste or powder form (see above).

Thus, the mixture provided in step (b) of the method of the present invention may be in the form of a mixture, in the preferred embodiment of the present invention in the form of a powder, or in the form of two, three, four or more liquids and/or gels, and /or paste-like and/or powdery pre-mixtures, which are present separately from each other, and which are mixed together before or during application or introduction onto/into the substrate to be treated in step (c).

An advantage of the present invention is that it is usually sufficient to perform steps (b) to (d) of the method of the present invention once to ensure satisfactory weed control.

However, according to another embodiment of the present invention, steps (b)-(d) or (b) and (c) can be repeated one, two, three or more times as necessary to ensure particularly effective bio-cementation of the substrate to be treated, and, thus particularly effective weed suppression.

Optionally, according to another embodiment of the present invention, one or more additional method steps may be performed prior to applying or introducing the mixture provided in step (b) onto/into the substrate to be treated (as defined in step (a) of the method of the present invention ), such as, for example, burning plants, in a preferred embodiment of the present invention, weeds located on/in the substrate, manual removal (weeding) of plants, in a preferred embodiment of the present invention, weeds located on/in the substrate, and/or treating plants, in a preferred embodiment of the present invention, weeds located on/in the substrate, with chemical reagents intended to control weeds. These steps can also be repeated one, two, three or more times, respectively.

A preferred embodiment of the present invention relates to the method described herein, wherein the formation of the biocement layer in step (d) of the method does not involve an exothermic reaction (as defined herein).

According to a preferred embodiment of the method of the present invention, the step of removing the substrate identified in step (a) on/in which plant growth, in the preferred embodiment of the present invention the growth of weeds, is to be prevented or reduced, is not necessary to prevent or reduce the growth plants and thus, in a preferred embodiment of the present invention, is not part of the method of the present invention, in particular since it is the thickening and/or hardening of the substrate during the formation of the biocement layer in step (d) of the method that causes the growth of plants to be prevented or reduced, in a preferred embodiment of the present invention, weed growth.

Moreover, within the framework of the method of the present invention described herein, in the preferred embodiment of the present invention, the step of compacting the substrate on/in which plant growth is to be reduced or prevented or the biocement layer formed to achieve the prevention or reduction of plant growth is not required. in a preferred embodiment of the present invention, weed growth, and thus, in a preferred embodiment of the present invention, this is not part of the method of the present invention.

What is stated herein with respect to the application of the present invention also applies to the method of the present invention as described herein, and vice versa. This applies in particular to (preferred) embodiments of the present invention which correspond to or can be derived from (preferred) embodiments of the method of the present invention, and vice versa.

The invention is described in more detail below by means of selected examples. Unless otherwise stated, all data refer to weight.

Blueprints:

fig. 1: suppression of weed growth through non-ureolytic biocementation using the bacterial strain B. pseudofirmus: Effect against monocotyledonous (Plantain meadow grass) and dicotyledonous (Plantain lanceolifolia) plants. Average weed growth rates over a 42-day documentation period with weekly monitoring of the control sample (top) in comparison with the sample treated with biocement mixture 1 (center). Visual representation (bottom) of weed growth when applied to the control sample (bottom left) compared to the sample treated with biocement mixture 1 (bottom right), after 42 days of growth.

Fig. 2: suppression of weed growth by non-ureolytic biocementation using bacterial strains A. crystallopoietes, B. cohnii, B. halodurans and B. pseudofirmus: Effect against monocotyledonous (Poa annual meadow grass) and dicotyledonous (Plantain lanceolifolia) plants. Average weed growth rates over a 42-day documentation period for weekly control measurements compared to samples treated with biocement mixture 1.

Fig. 3: use of ureolytic biocementation with L. sphaericus to suppress the growth of weeds in quartz sand: Effect against monocotyledonous (meadow grass annual) and dicotyledonous (plantain lanceolifolia) weeds. Average weed coverage rates (top) over a 42-day documentation period with weekly evaluation of the control sample compared to samples treated with Biocement Mix 2 and Biocement Mix 3. Visual representation (center) of weed growth when applied to the control sample (left in center) compared to samples treated with biocement mixture 2 (center) and biocement mixture 3 (right center), after 42 days of growth in laboratory conditions. Graphical representation of the thickening of biocement layers (bottom) by examining the average breaking force of the samples.

Fig. 4: use of ureolytic biocementation using L. sphaericus to suppress the growth of weeds in earthen soil: Action against monocotyledonous (annual meadow grass) and dicotyledonous (plantain lanceolate) weeds. Average weed coverage rates (top) over a 42-day documentation period with weekly evaluation of the control sample compared to samples treated with Biocement Mix 2 and Biocement Mix 3. Visual representation (center) of weed growth when applied to the control sample (left in center) compared to samples treated with biocement mixture 2 (center) and biocement mixture 3 (right center), after 42 days of growth in laboratory conditions. Graphical representation of the thickening of biocement layers (bottom) by examining the average breaking force of the samples.

Fig. 5: application of ureolytic biocementation using Sp. pasteurii to suppress the growth of weeds in quartz sand: Action against monocotyledonous (meadow grass annual) and dicotyledonous (plantain lanceolate) weeds. Average weed coverage rates (top) over a 42-day documentation period with weekly evaluation of the control sample compared to samples treated with Biocement Mix 4 and Biocement Mix 5. Visual representation (center) of weed growth when applied to the control sample (left in center) compared to samples treated with biocement mixture 4 (center) and biocement mixture 5 (right center), after 42 days of growth in laboratory conditions. Graphical representation of the thickening of biocement layers (bottom) by examining the average breaking force of the samples.

Fig. 6: application of ureolytic biocementation using Sp. pasteurii to suppress the growth of weeds in earthen soil: Action against monocotyledonous (meadow grass annual) and dicotyledonous (plantain lanceolate) weeds. Average weed coverage rates (top) over a 42-day documentation period with weekly evaluation of the control sample compared to samples treated with Biocement Mix 4 and Biocement Mix 5. Visual representation (center) of weed growth when applied to the control sample (left in center) compared to samples treated with biocement mixture 4 (center) and biocement mixture 5 (right center), after 42 days of growth in laboratory conditions. Graphical representation of the thickening of biocement layers (bottom) by examining the average breaking force of the samples.

Fig. 7: Application of biocementation to suppress the growth of weeds in open ground: Effect against ungerminated and freshly germinated weeds on agricultural land. Average coverage rates of water control (top) compared to sample treated with Biocement Mix 6 (center) over a 42-day documentation period with weekly assessments. Visual representation of weed growth when applied to the control sample (bottom left) compared to the sample treated with Biocement Mix 6 (bottom right), after 42 days of outdoor growth.

Fig. 8: the use of biocementation to suppress the growth of weeds in open ground: Effect against ungerminated and freshly sprouted weeds in sidewalk joints. Average coverage rates of water control (top) compared to sample treated with Biocement Mix 6 (center) over a 42-day documentation period with weekly assessments. Visual representation of weed growth when applied to the control sample (bottom left) compared to the sample treated with Biocement Mix 6 (bottom right), after 42 days of outdoor growth.

Examples:

Example 1. Non-ureolytic biocementation by B. pseudofirmus - suppression of the growth of monocotyledonous and dicotyledonous weeds.

Materials and methods:

The experiment was carried out in laboratory conditions in flower pots with a volume of 450 cm ³ . The area of application was 78.5 cm ² , respectively. A total of 6 samples were processed.

- 22 045836

The soil substrate involved in the experiment consisted of quartz sand with a grain size of 02 mm. The sand was washed and dried by the manufacturer and was used immediately. As a substrate with soil, 300 g of quartz sand was used per flower pot.

Before treatment, the quartz sand was free of weed growth and contained only remnants of endemic weed seeds or influx of seeds. However, this was not enough for the weeds to grow effectively. The weeds were inoculated using 0.2 g Plantago lanceolata and 0.1 g Poa annua per vessel, respectively. For this purpose, weed seeds were treated in the top layer of soil to a depth of 2-4 mm.

We used liquid biocement mixture 1, which consisted of the following components in the following concentrations:

20.0 g/l Yeast extract 0.2 mole calcium acetate 0.2 mole calcium lactate 6.0 g/l urea 5 x 10 ^l 8 cells/ml B. pseudofirmus

The mixture also contains trace elements and traces of salts and sugars, for example (<1 wt.%). In this environment, urea served primarily as a nitrogen source (rather than a carbonate source).

All components of the present mixture capable of biocementation, with the exception of bacteria strain B. pseudofirmus, were present in solid form. The bacteria were present in the form of a liquid culture in a culture medium known in the art, as described, for example, in the work of Yonkers H.M. et al., Custom Concrete Structures - Walraven & Stoelhorst (eds), 2008, Taylor & Francis Group, London, ISBN 978-0-415-47535-8, section 2.1, using 5 g/l yeast extract in context of the present invention. The solid components and bacteria in the liquid culture were mixed immediately before use, and the solid components were dissolved.

Biocement mixture 1 and water control were applied in triplicate to each of the study areas. Throughout, the number of applications per square meter was 5 liters per repetition. A pipette was used for application.

After applying biocement mixture 1, incubation occurred for 48 hours without irrigation. During this period, the minimum temperature was 14.2°C and the maximum was 25.2°C.

Weed growth was documented 42 days after application. The minimum and maximum temperatures during this period were 10.7°C and 34.0°C. The vessels were watered one to three times a week, depending on needs. Flower pots were exposed to natural light with a day and night rhythm.

Weed growth was documented weekly. Both the biocement layer (layer thickness, strength) and the so-called coverage rate were determined. Weed coverage rates were determined by manually visually assessing flower pots at the indicated time points. Coverage rate describes the percentage of area covered by weeds. From this, in turn, the Abbott degree of efficiency was calculated as follows:

Rate of effectiveness = (control sample coverage rate _{day xy} - product coverage rate day _xy )/control sample coverage rate _day xy.

To test the carbonate formation, 10 ml of biocement mixture 1 was incubated in air in a reaction vessel for 24 hours at room temperature. The precipitated granule was then obtained by centrifugation and drying. The dried granule was used for carbonate determination according to Scheibler.

Results:

Weed growth was almost completely reduced compared to the control sample (Fig. 1). The average coverage rate after 42 days was 2% in the treated area (Fig. 1, center) and 60% in the control area (Fig. 1, top). When treated with the above biocement mixture 1, a biocement layer was formed. Weed growth occurred mainly in areas where the biocement layer was damaged (for example, in drying cracks). The 42-day time period can be observed in FIG. 1 (top and center). Over time, the effect of biocementation in suppressing weeds is visible. In fig. 1 shows a direct comparison between a control sample (bottom left) and a sample to which the product of the present invention was applied (bottom right) after 42 days of growth. The final efficiency rate of the biocement product was 96.7%.

An advantage of the present invention is that the biocement mixture is as effective as many commercially available weed control agents (data not shown), thereby avoiding the various disadvantages associated with such weed control agents.

Qualitative analysis of the carbonate formation according to Scheibler showed a positive reaction to the biocement mixture. The control sample, on the other hand, did not show any carbonate formation (data not shown).

- 23 045836

Comparable effects on weed growth were also achieved when using slightly modified compositions of biocement mixture 1 containing calcium acetate, calcium lactate and/or calcium chloride in a concentration of 0.05-0.3 mol, respectively, and the total concentration of calcium in the mixture does not exceed 0.4 mol (data not shown). Changing the urea concentration (from 0.0 to 0.2 mol) or the amount of yeast extract (from 0.1 to 30 g/L) also showed good rates of effectiveness. Weed suppression depended on the applied concentrations of biocement mixture components, respectively (data not shown).

The entire experiment described above was carried out alternately with weed seeds that sprouted 24 hours before applying the biocement mixture. For this purpose, the biocement mixture was used 24 hours after the start of germination. The results obtained were comparable to those described in the present example, and almost complete reduction in weed growth was achieved with the mixture (data not shown).

Moreover, in the biocement mixture 1 described above, the bacterial strain B. pseudofirmus was replaced by the same cell number concentration of B. cohnii, B. halodurans or A. crystallopoietes, respectively, and the experiment was carried out as described above, respectively. B. cohnii and B. halodurans were present in the same culture medium as B. pseudofirmus (see above), and A. crystallopoietes was present in a known culture medium such as in Hamilton, R. W. et al., Journal of Bacteriology 1977, 129(2), 874-879 (see Materials and Methods, pp. 874-875). The results of weed suppression studies with these alternative biocement mixtures are shown in FIG. 2.

Example 2. Non-ureolytic biocementation by L. sphaericus - suppression of the growth of monocotyledonous and dicotyledonous weeds.

Materials and methods:

In the present experiment, two different biocement mixtures, each containing the same bacterial strain, were tested on two substrates with different soils.

The experiment was carried out in laboratory conditions in flower pots with a volume of 450 cm ³ . The area of application per vessel was 78.5 cm ² , respectively. A total of 9 flower pots per substrate with soil were treated with two different biocement mixtures (see below).

The first substrate with soil participating in the experiment consisted of quartz sand with a grain size of 0-2 mm. The quartz sand was washed and dried by the manufacturer and was used immediately. As a substrate with soil, 300 g of quartz sand was used per flower pot. In the next row, sifted earthen soil was used as the second substrate with soil. In this case, 250 g of earth soil per vessel was used to apply the mixture.

Both substrates and soil were free from weed growth before treatment. However, both soils contained minimal remnants of endemic weed seeds or seed influx. However, this was not enough for the weeds to grow effectively. The weeds were inoculated using 0.2 g Plantago lanceolata and 0.1 g Poa annua per vessel, respectively. For this purpose, the weeds were treated in the top layer of earthen soil to a depth of 2-4 mm.

Two different liquid biocement mixtures were used in the experiment.

Mixture 2 consisted of the following components in the following concentrations:

20.0 g/l Yeast extract 0.25 mole calcium chloride 18.0 g/l urea 4x 10 ^L 8 cells/ml L. sphaericus

The mixture also contained trace elements and traces of salts and sugars, for example (<1%). In this environment, urea served primarily as a carbonate source and secondarily as a nitrogen source.

To mixture 3, 50 ml/L Silica 8 (sol-acrylic silica dispersion) was added as an additive. The additive was used to achieve longer-term stability of the biocement layer.

The components of biocement mixtures 2 and 3 (without bacteria) were present in solid form, respectively. The bacteria were present in the form of a liquid culture in a culture medium known in the art, respectively, as described, for example, in Dick, J. et al., Biodegradation 2006, 17, 357-367 (see Materials and Methods, pp. 359). The solid components and bacteria in the liquid culture were mixed immediately before use, respectively, and the solid components were dissolved. Silicade 8 was present in liquid form and was only added to mixture 3.

Biocement mixtures 2 and 3, as well as a water control, were applied in triplicate next to each other to the two test soils. Throughout, the number of applications per square meter was 5 liters per repetition. A pipette was used for application.

After using biocement mixtures, incubation occurred for 48 hours without irrigation. During this period, the minimum temperature was 12.4°C and the maximum was 24.2°C.

Weed growth was documented 42 days after application. Minimum and

- 24 045836 the maximum temperature during this period was 9.7°C and 27.9°C. The vessels were watered one to three times a week, depending on needs. Flower pots were exposed to natural light with a day and night rhythm.

Weed growth was documented weekly. Both the biocement layer (layer thickness, strength) and the so-called coverage rate were determined. Weed coverage rates were determined by manually visually assessing flower pots at the indicated time points. Coverage rate describes the percentage of area covered by weeds. From this, in turn, the Abbott degree of efficiency was calculated as follows:

Effectiveness = (control sample coverage rate _{day xy} - product coverage rate _day xy)/control sample coverage rate _{day xy} .

To test the carbonate formation, 10 ml of biocement mixtures 2 and 3, respectively, were incubated in air in a reaction vessel for 24 hours at room temperature. The precipitated granule was then obtained by centrifugation and drying, respectively. The dried granules were used for carbonate determination according to Scheibler.

Results:

On quartz sand, weed growth was almost completely reduced compared to the control sample with both biocement mixtures 2 and 3 (Fig. 3). The average coverage rate after 42 days was 0% in the plot treated with biocement mixture 2, 0% in the plot treated with biocement mixture 3, and 31% in the control plot. As a result of both treatments (biocement mixtures 2 and 3), a biocement layer was formed. Weed growth occurred mainly in areas where the biocement layer was damaged (for example, in drying cracks). The 42-day time period can be observed in FIG. 3 (top). The effect of biocementation on weed suppression is shown in FIG. 3 (center), which shows a head-to-head comparison between the control sample (center left), Biocement Mix 2 (center), and Biocement Mix 3 (center right). The final efficiency of both biocement mixtures was 100%, respectively. After 42 days, the strength of the biocement layers was determined (as described above). The sample with biocementation with mixture 2 had a layer with an average destructive force of 4.3 N, however, this indicator is lower than that of the sample with biocementation with mixture 3 - 19.1 N (see Fig. 3 (below)). By incorporating the Silicad 8 additive into the biocement layer (via biocement mixture 3), increased resistance to environmental parameters and thus possibly greater efficiency could be achieved. In the control sample there was no biocement layer.

On earthen soil, weed growth was almost completely reduced compared to the control sample (Fig. 4). The average coverage rate after 42 days was 0% in the area treated with biocement mixture 2, 2% in the area treated with biocement mixture 3, and 50% in the control area. As a result of both treatments (biocement mixtures 2 and 3), a biocement layer was formed. Weed growth occurred mainly in areas where the biocement layer was damaged (for example, in drying cracks). The 42-day time period can be observed in FIG. 4 (top). The effect of biocementation on weed suppression is shown in FIG. 4 (center), which shows a head-to-head comparison between the control sample (center left), Biocement Mix 2 (center), and Biocement Mix 3 (center right). The final efficiency rates of the two biocement mixtures 2 and 3 were 100% and 96%, respectively. After 42 days, the strength of the resulting biocement layers was determined (as described above). The sample with biocementation with mixture 2 had a layer with an average destructive force of 20.5 N, however, this figure is lower than that of the sample with biocementation with mixture 3, which has a destructive force of 84.3 N. By including the additive Silicad 8 in the biocement layer (via biocement mixture 3) it would be possible to achieve increased resistance to environmental parameters and thus probably greater efficiency. In the control sample there was no biocement layer.

Qualitative analysis of carbonate formation according to Scheibler showed a positive reaction to biocement mixtures 2 and 3. Control samples did not show the presence of carbonate formation (data not shown).

Comparable effects on weed growth were also achieved using slightly modified formulations of biocement mixtures 2 and 3 containing calcium acetate, calcium lactate and/or calcium chloride at a concentration of 0.05-0.3 mol, respectively, and the total calcium concentration in the mixture does not exceed 0.4 mol (data not shown). Larger changes in urea concentration (eg, 0.1 to 1.0 mol) or yeast extract amount (eg, 0.1 to 30 g/L) also showed good efficacy rates. Weed suppression was dependent on the concentrations of components used in the respective biocement mixture, respectively (data not shown).

The entire experiment described above was carried out alternately with weed seeds that sprouted 24 hours before applying the appropriate biocement mixture. For this purpose, the appropriate biocement mixture was used 24 hours after the start of germination. The results obtained were comparable to the results described in this example and when using the appropriate mixture

- 25 045836 almost complete reduction in weed growth was achieved (data not shown).

Example 3. Non-ureolytic biocementation using Sp. pasteurii - suppression of the growth of monocotyledonous and dicotyledonous weeds.

Materials and methods:

In the present experiment, two different biocement mixtures, each containing the same bacterial strain, were tested on two substrates with different soils.

The experiment was carried out in laboratory conditions in flower pots with a volume of 450 cm ³ . The area of application was 78.5 cm ² , respectively. A total of 9 flower pots per substrate with soil were treated with two different biocement mixtures (see below). The area of application per vessel was 78.5 cm ² , respectively.

The first substrate with soil participating in the experiment consisted of quartz sand with a grain size of 0-2 mm. The quartz sand was washed and dried by the manufacturer and was used immediately. As a substrate with soil, 300 g of quartz sand was used per flower pot. In the next row, sifted earthen soil was used as the second substrate with soil. In this case, 250 g of earth soil per vessel was used to apply the mixture.

Both substrates and soil were free from weed growth before treatment. Both soils contained minimal traces of endemic weed seeds or seed influx. However, this was not enough for the weeds to grow effectively. The weeds were inoculated using 0.2 g Plantago lanceolata and 0.1 g Poa annua per vessel, respectively. For this purpose, weed seeds were treated in the top layer of soil to a depth of 2-4 mm.

Two different liquid biocement mixtures were used in the experiment.

Mixture 4 consisted of the following components in the following concentrations:

20.0 g/l Yeast extract 0.25 mole calcium chloride 18.0 g/l urea 4x 10 ^l 8 cells/ml Sp. pasteurii

The mixture also contained trace elements and traces of salts and sugars, for example (<1%). In this environment, urea served primarily as a carbonate source and secondarily as a nitrogen source.

To mixture 5, 50 ml/L Silica 8 (sol-acrylic silica dispersion) was added as an additive. The additive was used to achieve longer-term stability of the biocement layer.

Components of biocement mixtures 4 and 5 (without bacteria) were present in solid form, respectively. The bacteria were present in the form of a liquid culture in a culture medium known in the art, respectively, as described, for example, in Cuthbert, M. O. et al., Ecological Engineering 2012, 41, 32-40 (see section 2.2, page 33). The solid components and bacteria in the liquid culture were mixed immediately before mixing, respectively, and the solid components were dissolved. Silicade 8 was present in liquid form and was only added to mixture 5.

Biocement mixtures 4 and 5, as well as the water control, were applied in triplicate next to each other to the two test soils. Throughout, the number of applications per square meter was 5 liters per repetition. A pipette was used for application.

After using biocement mixtures, incubation occurred for 48 hours without irrigation. During this period, the minimum temperature was 12.4°C and the maximum was 24.2°C.

Weed growth was documented 42 days after application. The minimum and maximum temperatures during this period were 9.7°C and 27.9°C. The vessels were watered one to three times a week, depending on needs. Flower pots were exposed to natural light with a day and night rhythm.

Weed growth was documented weekly. Both the biocement layer (layer thickness, strength) and the so-called coverage rate were determined. Weed coverage rates were determined by manually visually assessing flower pots at the indicated time points. Coverage rate describes the percentage of area covered by weeds. From this, in turn, the Abbott degree of efficiency was calculated as follows:

Effectiveness = (control sample coverage rate _day xy - product coverage rate _day xy)/control sample coverage rate _day xy.

To test the carbonate formation, 10 ml of biocement mixtures 4 and 5, respectively, were incubated in air in a reaction vessel for 24 h at room temperature. The precipitated granule was then obtained by centrifugation and drying, respectively. The dried granules were used for carbonate determination according to Scheibler.

Results:

On quartz sand, weed growth was almost completely reduced compared to the control plot (Fig. 5). The average coverage rate after 42 days was 0% in the area treated with biocement mixture 4, 0% in the area treated with biocement mixture 5, and 40% in the control area. When treated with mixtures, a biocement layer was formed. Weed growth occurred mainly in areas where the biocement layer was damaged (for example, in drying cracks). The 42-day time period can be observed in FIG. 5 (top). The effect of biocementation on weed suppression is shown in FIG. 5 (center), which shows a head-to-head comparison between the control sample (center left), Biocement Mix 4 (center), and Biocement Mix 5 (center right). The final efficiency of both biocement mixtures was approximately 100%, respectively. After 42 days, the strength of the biocement layers was determined (as described above). The biocemented sample with mixture 4 had a layer with an average destructive force of 4.1 N, the biocemented sample with mixture 5 had an average destructive force of 19.3 N (see Fig. 5 (below)). By incorporating the Silicad 8 additive into the biocement layer (via biocement mixture 5), increased resistance to environmental parameters and thus possibly greater efficiency could be achieved. In the control sample there was no biocement layer.

On earthen soil, weed growth was almost completely reduced compared to the control sample (Fig. 6). The average coverage rate after 42 days was 0% in the area treated with biocement mixture 4, 0% in the area treated with biocement mixture 5, and 50% in the control area. When treated with mixtures, a biocement layer was formed. Weed growth occurred mainly in areas where the biocement layer was damaged (for example, in drying cracks). The 42-day time period can be observed in FIG. 6 (top). The effect of biocementation on weed suppression is shown in FIG. 6 (center), which shows a head-to-head comparison between the control sample (center left), Biocement Mix 4 (center), and Biocement Mix 5 (center right). The final efficiency rate of the two biocement mixtures was 100%, respectively. After 42 days, the strength of the resulting biocement layers was determined (as described above). The biocementation sample with mixture 4 had a layer with an average destructive force of 20.8 N, the biocementation sample with mixture 5 had an average destructive force of 66.8 N. By including the Silicad 8 additive in the biocement layer (via biocement mixture 5) Increased tolerance to environmental parameters and thus possibly greater efficiency could be achieved. In the control sample there was no biocement layer.

Qualitative analysis of the carbonate formation according to Scheibler showed a positive reaction to biocement mixtures 4 and 5, respectively. Control samples did not show the presence of carbonate formation (data not shown).

Comparable effects on weed growth were also achieved using slightly modified formulations of biocement mixtures 4 and 5 containing calcium acetate, calcium lactate and/or calcium chloride at a concentration of 0.05-0.3 mol, respectively, and the total calcium concentration in the mixture does not exceed 0.4 mol (data not shown). Larger changes in urea concentration (eg from 0.1 to 1.0 mol) also showed good efficacy rates. Weed suppression was dependent on the concentrations of components used in the respective biocement mixture, respectively (data not shown).

The entire experiment described above was carried out alternately with weed seeds that sprouted 24 hours before applying the appropriate biocement mixture. For this purpose, the appropriate biocement mixture was used 24 hours after the start of germination. The results obtained were comparable to those described in the present example, and almost complete reduction of weed growth was achieved with the appropriate mixture (data not shown).

Example 4. Open ground - suppression of weed growth on agricultural land and at sidewalk joints.

Materials and methods:

The experiment was conducted on agricultural land and on a cemented driveway. The application area was 6 cm ² , respectively.

The agricultural land soil substrate consisted of natural earthen soil. Before applying the mixture of the present invention (see below), the agricultural land was cleared of existing weeds by chemical treatment with glyphosate (approximately 6 months before the present experiment). After this pre-treatment, no plant residues remained on the surface.

The driveway bonding material consisted primarily of bonding gravel and bonding sand. Before use, these areas were mechanically cleared of existing weeds using a brush cutter. After this pre-treatment, no plant residues remained on the surface.

Both soils contained weed seeds, seed influx, and possibly fresh seedlings or plant debris present there. Artificial seeding of weeds was not carried out as there were sufficient endemic weeds in both sites.

For the experiment, liquid biocement mixture 6 was used, which consisted of the following components and concentrations:

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Claims (9)

18.0 g/l Urea 6 2.5 g/l lignosulfonate 5 x 10 ^L 8 cells/ml Sporosarcina pasteurii The mixture also contains trace elements and traces of salts, sugars and yeast extract, for example (<1%). The bacteria were present in the form of a liquid culture in the culture medium (see description in previous example 3). Urea and lignosulfonate were initially present in solid form. They were dissolved in water immediately before use and mixed with a liquid culture of bacteria. Biocement mixture 6 and water control were applied in triplicate to each of the two study sites, respectively. Throughout, the number of applications per square meter was 4 liters per repetition. For application, a standard watering can (volume 5 l) was used. After applying biocement mixture 6, incubation was carried out for 48 hours without rain or artificial irrigation. During this period, the minimum temperature was 5°C and the maximum was 25°C. Weed growth was documented 42 days after application. The minimum and maximum temperatures during this period were 5°C and 33°C. The total amount of precipitation during the documentation period was 91 mm (l/ ^m2 ). Thanks to the weather, no additional watering was required. Weed growth was documented weekly. Both the biocement layer (layer thickness, strength) and the so-called coverage rate were determined. Weed coverage rates were determined by manually visually assessing flower pots at the indicated time points. Coverage rate describes the percentage of area covered by weeds. From this, in turn, the Abbott degree of efficiency was calculated as follows: Effectiveness = (control sample coverage rate _{day xy} - product coverage rate _day xy)/control sample coverage rate _{day xy} . Results: On agricultural land, weed growth was significantly reduced compared to the control plot. The coverage rate after 42 days was 3.3% in the treated plots and 70.0% in the control plot. A biocement layer has formed. Weed growth occurred mainly in areas where the biocement layer was damaged (for example, in drying cracks). The 42-day time period can be observed in FIG. 7 (top, water control) and FIG. 7 (in the center, treatment with biocement mixture 6). In fig. Figure 7 (bottom) shows a direct comparison between the control sample and the application sample (marked with a marker, respectively). The final efficiency of biocement mixture 6 was 95.2%. Weed growth was also significantly reduced in the cemented driveway compared to the control plot. The coverage rate after 42 days was 3.7% in the treated plots and 40.0% in the control plot. A biocement layer has also formed here. The 42-day time period can be observed in FIG. 8 (top, water control) and FIG. 8 (in the center, treatment with biocement mixture 6). In fig. Figure 8 (bottom) shows a direct comparison between the control sample and the application sample (at joints). The final efficiency rate of the biocement product was 90.8%. An advantage of the present invention is that the biocement mixture is as effective as many commercially available weed control agents (data not shown), thereby avoiding the various disadvantages associated with such weed control agents. Comparable effects on outdoor weed growth were also shown with alternative mixture formulations additionally containing 0.1-0.3 mol CaCl ₂ (based on mixture 6) (data not shown). More significant changes in urea concentration (eg, from 1.0 to 0.15 mol) also showed good rates of effectiveness in suppressing weed growth (data not shown). CLAIM 1. The use of a mixture capable of biocementation as a means of preventing or reducing plant growth, in which the mixture contains or consists of the following components: (i) at least one organism and/or enzyme capable of forming carbonate and/or inducing and/or catalyzing carbonate formation, (ii) at least one substance for carbonate formation, (iii) at least one source of cations; and (iv) at least one additive selected from the group consisting of nutrients; (bio)polymers including polyhydroxybutyrate, polylactide, polybutylene succinate, polyacrylic acid, polymethacrylate, poly(2-hydroxyethyl methacrylate), polyvinyl alcohol, polyvinyl acetate, - 28 045836 polyvinylpyrrolidone, poly(2-ethyl-2-oxazoline), polystyrene, polyamide, copolymers, polyamic acids, cellulose and its derivatives, starch and its derivatives, lignins and their derivatives, pectins and their derivatives, natural glue, including Arabian gum, latex, rubber and their derivatives, chitin and its derivatives, chitosan and its derivatives, cyclodextrins and their derivatives, dextrins and their derivatives; hydrogelling agents including xanthan gum, alginates and agar-agar; cold-soluble and/or heat-soluble (vegetable) glue; calcium carbonates and mixtures containing calcium carbonates, including nacre, amorphous calcium carbonates, precipitated calcium carbonate, aragonite, calcite, vaterite and mixtures and derivatives thereof; extracellular polymeric substances (EPS) and polysaccharides, including microbial exopolysaccharides containing or consisting of maleic acid, acetic acid, lactic acid, lactose, sucrose, glucose, fructose and/or inulin; sources of protein, fibers and fibrous materials including casein, albumin, yeast extracts, peptones, cellulose fibers, wood fibers, wood pulp fibers; corn extract, lactose mother liquor, protein lysates, molasses, protein waste from yeast production, meat production, dairy and paper production; silicates and their derivatives; acrylates and their derivatives; wasserglass and binders similar to wasserglass; cement and cement additives including sand, lime and their derivatives, aluminum oxide, calcium oxide, calcium hydroxide, aluminum hydroxide, ash including fly ash and bone ash, microsilica, kaolin, bentonites, fillers including white lime (hydrate), limestone crushed sand and limestone powder; resins and epoxides; natural and chemical herbicides; fungicides; molluscicides; insecticides; water repellents and wax emulsions; emulsifiers; binders; thixotropic agents; crystallization nuclei and crystallization modifiers; fatty acid; minerals and trace elements; salts including phosphates and sulfates; rocks including pumice and shale powder; bacteria capable of forming polymers. 2. Use according to claim 1, wherein the mixture is present in liquid, gel, paste or powder form. 3. Use according to claim 1 or 2, wherein one or more or all of the above-mentioned organism(s) is/are selected from the group consisting of microorganisms, preferably selected/selected from the group consisting of microorganisms of the Firmicutes type, preferably the Bacilli class, preferably order Bacillales, preferably the families Planococcaceae or Bacillaceae, preferably the genera Sporosarcina, Lysinibacillus or Bacillus, preferably selected from the species Sporosarcina pasteurii, Sporosarcina ureae, Lysinibacillus sphaericus, Lysinibacillus fusiformis, Bacillus megaterium, Lysinibacillus sp., Bacillus pseudofirmus, Bacillus halodurans or Ba cillus cohnii; microorganisms of the phylum Proteobacteria, preferably of the classes Alphaproteobacteria, Gammaproteobacteria, Deltaproteobacteria or Epsilonproteobacteria, preferably of the orders Enterobacteriales, Myxococcales, Campylobacterales, Pseudomonadales or Caulobacterales, preferably of the families Enterobacteriaceae, Myxococcaceae, Helicobacteraceae, Pseudomonadaceae or Caulobacteraceae, preferably the genera Proteus, Myxococcus, Helicobacter, Pseudomonas or Brevundimonas, preferably selected from the species Proteus vulgaris, Proteus mirabilis, Myxococcus xanthus, Helicobacter pylori, Pseudomonas aeruginosa or Brevundimonas diminuta; microorganisms of the phylum Actinobacteria, preferably the class Actinobacteria, preferably the order Actinomycetales, preferably the families Brevibacteriaceae or Micrococcineae, preferably the genera Brevibacterium or Micrococcaceae, preferably selected from the species Brevibacterium lines or Arthrobacter crystallopoietes; microorganisms of the phylum Cyanobacteria, preferably the class Cyanobacteria, preferably the order Synechococcales, preferably the family Synechococcaceae, preferably the genus Synechococcus, preferably the species Synechococcus. 4. Use according to any of the preceding claims, wherein one, more or all of the above enzymes are/are selected from the group consisting of ureases, asparaginases, carbonic anhydrases and metabolic enzymes. 5. Use according to any of the previous claims, wherein one, more or all of the above-mentioned carbonate-forming substances are selected from the group consisting of urea and its salts, organic acids such as lactic acid and its salts, preferably carboxylates and esters thereof, gluconic acid and its salts, preferably carboxylates and their esters, acetic acid and its salts, preferably carboxylates and their esters, formic acid and its salts, preferably carboxylates and their esters, peptides preferably containing asparagine, glutamine and/or glutamic acid, amino acids , preferably asparagine, glutamine and glutamic acid and its salts, preferably carboxylates and their esters, plant and animal complex substrates, in particular peptone, yeast extract, meat extract, nutrient broth and casaminic acid, corn extract, lactose stock solution, protein lysates , preferably from peas, meat or tomatoes, from carbon dioxide and methane. 6. Use according to any of the preceding claims, wherein one, more or all of the above cation sources are/are selected from the group consisting of organic and inorganic calcium salts, preferably calcium nitrate, calcium acetate, calcium lactate and calcium chloride, magnesium salts, manganese salts , zinc salts, cobalt salts, nickel salts, copper salts, lead salts, salts - 29 045836 iron, cadmium salts, polymers, preferably cationic polymers, heavy metal cations, light metal cations, radioactive cations and mixtures thereof. 7. Application according to any of the previous paragraphs, in which the plant is selected from the group including dicotyledonous plants of the following genera: Abutilon, Snit, Kokorysh, Shchiritsa, Ambrosia, Ankhusa, Anagallis, Anode, Antemis, Nevzrachnitsa, Arabidopsis, Quinoa, Crescent, Daisy, Sequence, Sverbiga, Shepherd's purse, Thistle, Cassia, Cornflower, Goosefoot, Chrysanthemum, Thistle, Hemlock, Conise, Consolidum, Convolvulus, Datura, Descurainia, Desmodium, Emex, Horsetail, Erigeron, Erodium, Jaundice, Euphorbia, Furmaria, Pikulnik, Galinzoga , Bedstraw, Geranium, Hogweed, Hibiscus, Morning Glory, Kochia, Lathyrus, Lepidium, Lithospermum, Linaria, Lindernia, Crookedflower, Mallow, Matricaria, Mint, Woodweed, Mollugo, Forget-me-not, Papaver, Farbitis, Plantain, Polygonum, Purslane , Ranunculus, Radish, Zherushnik, Rotala, Sorrel, Solyanka, Rasson, Sesbania, Sida, Sinapis, Gooseberry, Solanum, Sow thistle, Sphenoclea, Stachys, Chickweed, Dandelion, Jarutka, Clover, Coltsfoot, Nettle, Veronica, Viola , cocklebur; dicotyledonous plants of the following genera: Peanut, Beetroot, Brassica, Cucumis, Pumpkin, Helianthus, Carrot, Wisteria, Cotton, Morning Glory, Lettuce, Flax, Tomato, Tobacco, Beans, Peas, Solanum, Peas; monocotyledonous plants of the following genera: Aegilops, Zhitnyak, Bentgrass , Alopecurus, Metlitsa, Oats, Brachiaria, Bonfire, Cenchrus, Commelina, Pigwort, Cyperus, Dactyloctenium, Crabgrass, Barnyardgrass, Eleocharis, Eleusina, Eragrostis, Woolgrass, Fescue, Fimbristylis, Heteranthera, Imperata, Murraya, Juncus, Leptochloa, Tares, Monochoria , Panicum, Paspalum, Canary grass, Timothy grass, Bluegrass, Rothbellia, Sagittaria, Scirpus, Setaria, Sorghum; and monocotyledons of the following genera: Allium, Pineapple, Asparagus, Oats, Barley, Rice, Panicum, Sugarcane, Rye, Sorghum, Triticale, Wheat, Corn; mosses of the liverwort class, hornworts, mosses (Bryopsida); preferably, the growth of at least two, three, four, five, six, seven, eight, nine, ten, more than ten or all of these plants is prevented or reduced. 8. A mixture capable of biocementing a substrate, characterized in that it contains or consists of: (i) at least one organism and/or enzyme capable of forming carbonate, inducing carbonate formation and/or catalyzing carbonate formation; (ii) at least one carbonate-forming substance; (iii) at least one cation source; and (iv) at least one additive selected from nutrients; (bio)polymers including polyhydroxybutyrate, polylactide, polyacrylic acid, polymethacrylate, poly(2hydroxyethyl methacrylate), polyvinyl alcohol, polyvinyl acetate, polyvinylpyrrolidone, poly(2-ethyl2-oxazoline), polystyrene, copolymers, polyamic acids, cellulose and its derivatives, starch and its derivatives, lignins and their derivatives, pectins and their derivatives, natural adhesives including gum arabic, latex, chitin and its derivatives, chitosan and its derivatives, cyclodextrins and their derivatives, dextrins and their derivatives; hydrogel formers, including xanthan gum, alginates; cold-soluble and/or heat-soluble (vegetable) adhesives; calcium carbonates, including mother of pearl, amorphous calcium carbonates, precipitated calcium carbonates, aragonite, calcite, vaterite and mixtures and derivatives thereof; extracellular polymeric substances and polysaccharides, including microbial exopolysaccharides containing or consisting of maleic acid, acetic acid, lactic acid, lactose, sucrose, glucose, fructose and/or inulin; polysaccharide monomers including lactose, sucrose, glucose, fructose and/or inulin; sources of protein, fibers and fibrous materials including casein, albumin, yeast extracts, peptones, cellulose fibers, wood fibers, wood pulp fibers; casein monomers; albumin monomers; corn extract, lactose mother liquor, protein lysates, molasses, protein waste from yeast production, meat production, dairy and paper production; silicates and their derivatives; acrylates and their derivatives; wasserglass and binders similar to wasserglass; cement additives, aluminum oxide, calcium oxide, calcium hydroxide, aluminum hydroxide, ash, including fly ash and bone ash, microsilica, kaolins, bentonites; resins and epoxides; natural and chemical herbicides; fungicides; molluscicides; insecticides; water repellents and wax emulsions; emulsifiers; thixotropic agents; crystallization nuclei and crystallization modifiers; fatty acids; microelements; salts including phosphates and sulfates; bacteria capable of forming polymers; wherein said at least one additive is present in an amount that provides additional thickening and/or hardening of the substrate, and/or increasing its stability. 9. A method of preventing or reducing the growth of plants on/in a substrate, consisting or containing the following steps: (a) identifying the substrate to be treated on/in which plant growth is to be prevented or reduced, (b) providing the mixture according to claim 9. (c) applying and/or introducing the mixture provided in step (b) onto/into the substrate to be treated in an amount sufficient to ensure bio-cementation, and (d) forming a bio-cement layer in such a way as to prevent or reduce the growth of dis- -